3-(heteroaryl)alanine derivatives-inhibitors of leukocyte adhesion mediated by vla-4

ABSTRACT

Disclosed are certain 3-(heteroaryl)alanine derivatives which bind VLA-4 and inhibit leukocyte adhesion mediated by VLA-4. Such compounds are useful in the treatment of inflammatory diseases in a mammalian patient, e.g., human, such as asthma, Alzheimer&#39;s disease, atherosclerosis, AIDS dementia, diabetes, inflammatory bowel disease, rheumatoid arthritis, tissue transplantation, tumor metastasis and myocardial ischemia. The compounds can also be administered for the treatment of inflammatory brain diseases such as multiple sclerosis.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application Ser.No. 60/220,131, filed Jul. 21, 2000, which is incorporated by referencein its entirety.

BACKGROUND OF THE INVENTION Field of the Invention

This invention relates to certain 3-(heteroaryl)alanine derivativeswhich inhibit leukocyte adhesion and, in particular, leukocyte adhesionmediated by VLA-4.

REFERENCES

The following publications, patents and patent applications are cited inthis application as superscript numbers:

-   -   ¹ Hemler and Takada, European Patent Application Publication No.        330,506, published Aug. 30, 1989    -   ² Elices, et al., Cell, 60:577-584 (1990)    -   ³ Springer, Nature, 346:425-434 (1990)    -   ⁴ Osborn, Cell, 62:3-6 (1990)    -   ⁵ Vedder, et al., Surgery, 106:509 (1989)    -   ⁶ Pretolani, et al., J. Exp. Med., 180:795 (1994)    -   ⁷ Abraham, et al., J. Clin. Invest., 93:776 (1994)    -   ⁸ Mulligan, et al., J. Immunology, 150:2407 (1993)    -   ⁹ Cybulsky, et al., Science, 251:788 (1991)    -   ¹⁰ Li, et al., Arterioscler. Thromb., 13:197 (1993)    -   ¹¹ Sasseville, et al., Am. J. Path., 144:27 (1994)    -   ¹² Yang, et al., Proc. Nat. Acad. Science (USA), 90:10494 (1993)    -   ¹³ Burkly, et al., Diabetes, 43:529 (1994)    -   ¹⁴ Baron, et al., J. Clin. Invest., 93:1700 (1994)    -   ¹⁵ Hamann, et al., J. Immunology, 152:3238 (1994)    -   ¹⁶ Yednock, et al., Nature, 356:63 (1992)    -   ¹⁷ Baron, et al., J. Exp. Med., 177:57 (1993)    -   ¹⁸ van Dinther-Janssen, et al., J. Immunology, 147:4207 (1991)    -   ¹⁹ van Dinther-Janssen, et al., Annals. Rheumatic Dis., 52:672        (1993)    -   ²⁰ Elices, et al., J. Clin. Invest., 93:405 (1994)    -   ²¹ Postigo, et al., J. Clin. Invest., 89:1445 (1991)    -   ²² Paul, et al., Transpl. Proceed., 25:813 (1993)    -   ²³ Okarhara, et al., Can. Res., 54:3233 (1994)    -   ²⁴ Paavonen, et al., Int. J. Can., 58:298 (1994)    -   ²⁵ Schadendorf, et al., J. Path., 170:429 (1993)    -   ²⁶ Bao, et al., Diff., 52:239 (1993)    -   ²⁷ Lauri, et al., British J. Cancer, 68:862 (1993)    -   ²⁸ Kawaguchi, et al., Japanese J. Cancer Res., 83:1304 (1992)    -   ²⁹ Kogan, et al., U.S. Pat. No. 5,510,332, issued Apr. 23, 1996    -   ³⁰ International Patent Appl. Publication No. WO 96/01644

All of the above publications, patents and patent applications areherein incorporated by reference in their entirety to the same extent asif each individual publication, patent or patent application wasspecifically and individually indicated to be incorporated by referencein its entirety.

State of the Art

VLA-4 (also referred to as α₄β₁integrin and CD49d/CD29), firstidentified by Hemler and Takada¹ is a member of the β1 integrin familyof cell surface receptors, each of which comprises two subunits, an αchain and a β chain. VLA-4 contains an α4 chain and a β1 chain. Thereare at least nine β1 integrins, all sharing the same β1 chain and eachhaving a distinct a chain. These nine receptors all bind a differentcomplement of the various cell matrix molecules, such as fibronectin,laminin, and collagen. VLA-4, for example, binds to fibronectin. VLA-4also binds non-matrix molecules that are expressed by endothelial andother cells. These non-matrix molecules include VCAM-1, which isexpressed on cytokine-activated human umbilical vein endothelial cellsin culture. Distinct epitopes of VLA-4 are responsible for thefibronectin and VCAM-1 binding activities and each activity has beenshown to be inhibited independently.²

Intercellular adhesion mediated by VLA-4 and other cell surfacereceptors is associated with a number of inflammatory responses. At thesite of an injury or other inflammatory stimulus, activated vascularendothelial cells express molecules that are adhesive for leukocytes.The mechanics of leukocyte adhesion to endothelial cells involves, inpart, the recognition and binding of cell surface receptors onleukocytes to the corresponding cell surface molecules on endothelialcells. Once bound, the leukocytes migrate across the blood vessel wallto enter the injured site and release chemical mediators to combatinfection. For reviews of adhesion receptors of the immune system, see,for example, Springer³ and Osborn⁴.

Inflammatory brain disorders, such as experimental autoimmuneencephalomyelitis (EAE), multiple sclerosis (MS) and meningitis, areexamples of central nervous system disorders in which theendothelium/leukocyte adhesion mechanism results in destruction tootherwise healthy brain tissue. Large numbers of leukocytes migrateacross the blood brain barrier (BBB) in subjects with these inflammatorydiseases. The leukocytes release toxic mediators that cause extensivetissue damage resulting in impaired nerve conduction and paralysis.

In other organ systems, tissue damage also occurs via an adhesionmechanism resulting in migration or activation of leukocytes. Forexample, it has been shown that the initial insult following myocardialischemia to heart tissue can be further complicated by leukocyte entryto the injured tissue causing still further insult (Vedder et al.⁵).Other inflammatory or medical conditions mediated by an adhesionmechanism include, by way of example, asthma⁶⁻⁸, Alzheimer's disease,atherosclerosis⁹⁻¹⁰, AIDS dementia¹¹, diabetes¹²⁻¹⁴ (including acutejuvenile onset diabetes), inflammatory bowel disease¹⁵ (includingulcerative colitis and Crohn's disease), multiple sclerosis¹⁶⁻¹⁷,rheumatoid arthritis¹⁸⁻²¹, tissue transplantation²², tumormetastasis²³⁻²⁸, meningitis, encephalitis, stroke, and other cerebraltraumas, nephritis, retinitis, atopic dermatitis, psoriasis, myocardialischemia and acute leukocyte-mediated lung injury such as that whichoccurs in adult respiratory distress syndrome.

In view of the above, assays for determining the VLA-4 level in abiological sample containing VLA-4 would be useful, for example, todiagnosis VLA-4 mediated conditions. Additionally, despite theseadvances in the understanding of leukocyte adhesion, the art has onlyrecently addressed the use of inhibitors of adhesion in the treatment ofinflammatory brain diseases and other inflammatory conditions^(29,30).The present invention addresses these and other needs.

SUMMARY OF THE INVENTION

This invention provides certain 3-(heteroaryl)alanine derivatives whichbind to VLA-4. Such compounds can be used, for example, to assay for thepresence of VLA-4 in a sample and in pharmaceutical compositions toinhibit cellular adhesion mediated by VLA-4, for example, binding ofVCAM-1 to VLA-4. The compounds of this invention have a binding affinityto VLA-4 as expressed by an IC₅₀ of about 15 μM or less (as measuredusing the procedures described in Example A below).

In particular, this invention is based on the discovery that thepresence of a nitrogen containing heteroaryl group at the 3-position ofthese alanine derivatives provides significant advantages vis-a-vis arylor other heteroaryl groups at this position in the molecule.

Accordingly, in one of its composition aspects, this invention isdirected to a compound of Formula (I):

wherein:

A is selected from the group consisting of aryl, heteroaryl, cycloalkyl,and heterocyclic group wherein said aryl, heteroaryl, cycloalkyl, orheterocyclic group is optionally substituted, on any ring atom capableof substitution, with 1-3 substituents selected from the groupconsisting of alkyl, substituted alkyl, alkoxy, substituted alkoxy,acyl, acylamino, thiocarbonylamino, acyloxy, amino, substituted amino,amidino, alkyl amidino, thioamidino, aminoacyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aryl, substituted aryl,aryloxy, substituted aryloxy, aryloxyaryl, substituted aryloxyaryl,cyano, halogen, hydroxyl, nitro, oxo, carboxyl, cycloalkyl, substitutedcycloalkyl, guanidino, guanidinosulfone, thiol, thioalkyl, substitutedthioalkyl, thioaryl, substituted thioaryl, thiocycloalkyl, substitutedthiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,thioheterocyclic, substituted thioheterocyclic, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —OS(O)₂-alkyl, —OS(O)₂-substituted alkyl,—OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where each R isindependently hydrogen or alkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substitutedalkyl, —NRS(O)₂-aryl, —NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl,—NRS(O)₂-substituted heteroaryl, —NRS(O)₂-heterocyclic,—NRS(O)₂-substituted heterocyclic, —NRS(O)₂—NR-alkyl,—NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl, —NRS(O)₂—NR-substitutedaryl, —NRS(O)₂—NR-heteroaryl, —NRS(O)₂—NR-substituted heteroaryl,—NRS(O)₂—NR-heterocyclic, —NRS(O)₂—NR-substituted heterocyclic where Ris hydrogen or alkyl, —N[S(O)₂—R′]₂ and —N[S(O)₂—NR′]₂ where each R′ isindependently selected from the group consisting of alkyl, substitutedalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic;

HetAr is a nitrogen containing heteroaryl or a nitrogen containingsubstituted heteroaryl group;

Alk is an alkylene group of 1 to 4 carbons;

m is 0 or 1;

R¹ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic and substitutedheterocyclic;

X is selected from the group consisting of hydroxyl, alkoxy, substitutedalkoxy, alkenoxy, substituted alkenoxy, cycloalkoxy, substitutedcycloalkoxy, cycloalkenoxy, substituted cycloalkenoxy, aryloxy,substituted aryloxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy and —NR″R″ where each R″ isindependently selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, substitutedcycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic;

and enantiomers, diasteromers and pharmaceutically acceptable saltsthereof;

and further wherein the compound of Formula (I) has a binding affinityto VLA-4 as expressed by an IC₅₀ of about 15 μM or less.

In a preferred embodiment, Alk is preferably 1 carbon and m ispreferably 1.

In a preferred embodiment, HetAr in the above compounds is a nitrogencontaining substituted heteroaryl group, preferably a nitrogencontaining heteroaryl group that is substituted with a substituentselected from the group consisting of acyl, acylamino, acyloxy,aminoacyl, aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy,oxycarbonylamino, oxythiocarbonylamino, thioamidino, thiocarbonylamino,aminosulfonylamino, aminosulfonyloxy, aminosulfonyl, oxysulfonylaminooxysulfonyl, aryl and substituted aryl.

Within this group a more preferred group is wherein the nitrogencontaining heteroaryl group is substituted with a substituent of formula—O-Z-NR¹¹R^(11′) or —O-Z-R¹² wherein R¹¹ and R^(11′) are independentlyselected from the group consisting of hydrogen, alkyl, substitutedalkyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, heterocyclic, substituted heterocyclic, and where R¹¹ andR^(11′) are joined to form a heterocycle or a substituted heterocycle,R¹² is selected from the group consisting of heterocycle and substitutedheterocycle, and Z is selected from the group consisting of —C(O)— and—SO₂—. More preferably, the nitrogen containing heteroaryl group issubstituted with a group of formula —OC(O)NR¹¹R^(11′), wherein R¹¹ andR^(11′) are as defined herein. Even more preferably —OC(O)NR¹¹R^(11′)wherein R¹¹ and R^(11′) are independently selected from the groupconsisting of alkyl or R¹¹ and R^(11′) are joined to form a heterocycleor a substituted heterocycle, most preferably ′OC(O)N(CH₃)₂.

Another more preferred group is wherein the nitrogen containingheteroaryl group is substituted with an aryl or substituted arylsubstituent. More preferably, the heteroaryl group is substituted withdialkoxyphenyl.

In yet another preferred embodiment, A in the above compounds isheteroaryl optionally substituted with 1 to 3 substituents selected fromthe group consisting of alkyl, substituted alkyl, alkoxy, substitutedalkoxy, amino, substituted amino, cycloalkyl, substituted cycloalkyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic and halogen. Preferably, A isselected from the group consisting of 1-oxo-1,2,5-thiadiazole,1,1-dioxo-1,2,5-thiadiazole, pyridazine, pyrimidine or pyrazine; morepreferably, pyrimidine or pyrazine; wherein the pyridazine, pyrimidineor pyrazine ring is optionally substituted with 1 to 3 substituentsselected from the group consisting of alkyl, substituted alkyl, alkoxy,substituted alkoxy, amino, substituted amino, cycloalkyl, substitutedcycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic and halogen.

In yet another preferred embodiment, R¹ is hydrogen, and X is hydroxyl.

In still another preferred embodiment, this invention is directed tocompounds of Formula IIa, IIb, IIc, IId, or IIe:

wherein:

HetAr is a nitrogen containing heteroaryl group substituted at anyposition capable of substitution with a single substitution selectedfrom the group consisting of acyl, acylamino, acyloxy, aminoacyl,aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy,oxycarbonylamino, oxythiocarbonylamino, thioamidino, thiocarbonylamino,aminosulfonylamino, aminosulfonyloxy, aminosulfonyl, oxysulfonylamino,aryl, substituted aryl, and oxysulfonyl;

R⁵ is selected from the group consisting of alkyl, substituted alkyl,alkenyl, substituted alkenyl, aryl, substituted aryl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl,heterocyclic, substituted heterocylic, heteroaryl and substitutedheteroaryl;

R⁶ is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, heterocyclic, substituted heterocyclic, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, and —SO₂R¹⁰ where R¹⁰ isselected from the group consisting of alkyl, substituted alkyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, heterocyclic, substituted heterocyclic, aryl, substitutedaryl, heteroaryl, substituted heteroaryl;

R⁷ and R⁸ are independently selected from the group consisting ofhydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic and halogen;

R¹⁶ and R¹⁷ are independently selected from the group consisting ofhydrogen, alkyl, substituted alkyl, alkoxy, substituted alkoxy, amino,substituted amino, cycloalkyl, substituted cycloalkyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic and halogen; and

R¹⁸ is selected from the group consisting of alkyl, substituted alkyl,alkoxy, substituted alkoxy, amino, substituted amino, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic;

R²⁰ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl, substitutedcycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic and halogen;

R²¹ is selected from the group consisting of alkyl, substituted alkyl,alkoxy, substituted alkoxy, amino, substituted amino, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heterocyclic andsubstituted heterocyclic;

b is 1 or 2; and

X is hydroxyl; and

and enantiomers, diastereomers and pharmaceutically acceptable saltsthereof.

Preferably, the compound is selected from Formula IIc, IId or IIe.

In the above compounds II(a-e), HetAr is preferably:

a) a nitrogen containing heteroaryl ring which is substituted with agroup of formula —O-Z-NR¹¹R^(11′) or —O-Z-R¹² wherein R¹¹ and R^(11′)are independently selected from the group consisting of hydrogen, alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, heterocyclic, substituted heterocyclic, andwhere R¹¹ and R^(11′) are joined to form a heterocycle or a substitutedheterocycle, R¹² is selected from the group consisting of heterocycleand substituted heterocycle, and Z is selected from the group consistingof —C(O)— and —SO₂—. More preferably, the nitrogen containing heteroarylring is substituted with a group of formula —OC(O)NR¹¹R^(11′), whereinR¹¹ and R^(11′) are as defined herein. Even more preferably—OC(O)NR¹¹R^(11′) wherein R¹¹ and R^(11′) are independently selectedfrom the group consisting of alkyl or R¹¹ and R^(11′) are joined to forma heterocycle or a substituted heterocycle, most preferably—OC(O)N(CH₃)₂; or

b) a nitrogen containing heteroaryl ring which is substituted with anaryl or substituted aryl group. More preferably, wherein the heteroarylring is substituted with dialkoxyphenyl.

Another preferred group of compounds are those wherein R⁵ is selectedfrom the group consisting of alkyl, substituted alkyl, aryl, substitutedaryl, heterocyclic, substituted heterocylic, heteroaryl and substitutedheteroaryl. Even more preferably R⁵ is selected from the groupconsisting of 4-methylphenyl, methyl, benzyl, n-butyl, n-hexyl,4-chlorophenyl, 1-naphthyl, 2-naphthyl, 4-methoxyphenyl, phenyl,2,4,6-trimethylphenyl, 2-(methoxycarbonyl)phenyl, 2-carboxyphenyl,3,5-dichlorophenyl, 4-trifluoromethylphenyl, 3,4-dichlorophenyl,3,4-dimethoxyphenyl, 4-(CH₃C(O)NH-)phenyl, 4-trifluoromethoxyphenyl,4-cyanophenyl, isopropyl, 3,5-di-(trifluoromethyl)phenyl,4-t-butylphenyl, 4-t-butoxyphenyl, 4-nitrophenyl, 2-thienyl,1-N-methyl-3-methyl-5-chloropyrazol-4-yl, phenethyl,1-N-methylimidazol-4-yl, 4-bromophenyl, 4-amidinophenyl,4-methylamidinophenyl, 4-[CH₃SC(═NH)]phenyl, 5-chloro-2-thienyl,2,5-dichloro-4-thienyl, 1-N-methyl-4-pyrazolyl, 2-thiazolyl,5-methyl-1,3,4-thiadiazol-2-yl, 4-[H₂NC(S)]phenyl, 4-aminophenyl,4-fluorophenyl, 2-fluorophenyl, 3-fluorophenyl, 3,5-difluorophenyl,pyridin-3-yl, pyrimidin-2-yl, 4-(3′-dimethylamino-n-propoxy)-phenyl, and1-methylpyrazol-4-yl;

R¹⁶ is substituted amino;

R⁶, R¹⁷ and/or R²⁰ are hydrogen; and

R¹⁸ and/or R²¹ are alkyl, substituted alkyl, aryl, or substituted aryl.

In a second aspect, this invention provides pharmaceutical compositionscomprising a pharmaceutically acceptable carrier and a therapeuticallyeffective amount of the compounds defined herein.

In a third aspect, this invention is directed to a method for treating adisease mediated by VLA-4 in a patient, which method comprisesadministering a pharmaceutical composition comprising a pharmaceuticallyacceptable carrier and a therapeutically effective amount of compoundsdefined herein.

The compounds and pharmaceutical compositions of this invention areuseful for treating disease conditions mediated by VLA-4 or leucocyteadhesion. Such disease conditions include, by way of example, asthma,Alzheimer's disease, atherosclerosis, AIDS dementia, diabetes (includingacute juvenile onset diabetes), inflammatory bowel disease (includingulcerative colitis and Crohn's disease), multiple sclerosis, rheumatoidarthritis, tissue transplantation, tumor metastasis, meningitis,encephalitis, stroke, and other cerebral traumas, nephritis, retinitis,atopic dermatitis, psoriasis, myocardial ischemia and acuteleukocyte-mediated lung injury such as that which occurs in adultrespiratory distress syndrome.

Other disease conditions include, but are not limited to, inflammatoryconditions such as erythema nodosum, allergic conjunctivitis, opticneuritis, uveitis, allergic rhinitis, Ankylosing spondylitis, psoriaticarthritis, vasculitis, Reiter's syndrome, systemic lupus erythematosus,progressive systemic sclerosis, polymyositis, dermatomyositis, Wegner'sgranulomatosis, aortitis, sarcoidosis, lymphocytopenia, temporalarteritis, pericarditis, myocarditis, congestive heart failure,polyarteritis nodosa, hypersensitivity syndromes, allergy,hypereosinophilic syndromes, Churg-Strauss syndrome, chronic obstructivepulmonary disease, hypersensitivity pneumonitis, chronic activehepatitis, interstitial cystitis, autoimmune endocrine failure, primarybiliary cirrhosis, autoimmune aplastic anemia, chronic persistenthepatitis and thyroiditis.

In a preferred embodiment, the disease condition mediated by VLA-4 is aninflammatory disease.

In the above compounds, when X is other than —OH or pharmaceutical saltsthereof, X is preferably a substituent which will convert (e.g.,hydrolyze, metabolize, etc.) in vivo to a compound where X is —OH or asalt thereof. Accordingly, suitable X groups are any art recognizedpharmaceutically acceptable groups which will hydrolyze or otherwiseconvert in vivo to a hydroxyl group or a salt thereof including, by wayof example, esters (X is alkoxy, substituted alkoxy, cycloalkoxy,substituted cycloalkoxy, alkenoxy, substituted alkenoxy, cycloalkenoxy,substituted cycloalkenoxy, aryloxy, substituted aryloxy, heteroaryloxy,substituted heteroaryloxy, heterocyclooxy, substituted heterocyclooxy,and the like).

This invention also provides methods for binding VLA-4 in a biologicalsample which method comprises contacting the biological sample with acompound of this invention under conditions wherein said compound bindsto VLA-4.

The pharmaceutical compositions may be used to treat disease conditionsmediated by VLA-4 or leucocyte adhesion. Such disease conditionsinclude, by way of example, asthma, Alzheimer's disease,atherosclerosis, AIDS dementia, diabetes (including acute juvenile onsetdiabetes), inflammatory bowel disease (including ulcerative colitis andCrohn's disease), multiple sclerosis, rheumatoid arthritis, tissuetransplantation, tumor metastasis, meningitis, encephalitis, stroke, andother cerebral traumas, nephritis, retinitis, atopic dermatitis,psoriasis, myocardial ischemia and acute leukocyte-mediated lung injurysuch as that which occurs in adult respiratory distress syndrome.

Other disease conditions include, but are not limited to, inflammatoryconditions such as erythema nodosum, allergic conjunctivitis, opticneuritis, uveitis, allergic rhinitis, ankylosing spondylitis, psoriaticarthritis, vasculitis, Reiter's syndrome, systemic lupus erythematosus,progressive systemic sclerosis, polymyositis, dermatomyositis, Wegner'sgranulomatosis, aortitis, sarcoidosis, lymphocytopenia, temporalarteritis, pericarditis, myocarditis, congestive heart failure,polyarteritis nodosa, hypersensitivity syndromes, allergy,hypereosinophilic syndromes, Churg-Strauss syndrome, chronic obstructivepulmonary disease, hypersensitivity pneumonitis, chronic activehepatitis, interstitial cystitis, autoimmune endocrine failure, primarybiliary cirrhosis, autoimmune aplastic anemia, chronic persistenthepatitis and thyroiditis.

Preferred compounds of this invention include those set forth in Table Ibelow:

TABLE I

Cpd # A HetAr 1 [2-N(CH₃)₂-5-CF₃CH₂]pyrimi- 2-(2,6-di-MeOPh)pyridin-5-yldin-4-yl 2 [2-N(CH₃)₂-5-CF₃CH₂]pyrimi- 5-(2,6-di-MeOPh)pyridin-2-yldin-4-yl 3 [2-N(CH₃)₂-5-CF₃CH₂]pyrimi- 3-(2,6-di-MeOPh)pyrida- din-4-ylzin-6-yl 4 [2-N(CH₃)₂-5-CF₃CH₂]pyrimi- 5-(2,6-di-MeOPh)pyrimi- din-4-yldin-5-yl 5 [2-N(CH₃)₂-5-CF₃CH₂]pyrimi- 2-(2,6-di-MeOPh)pyrimi- din-4-yldin-5-yl 6 [2-N(CH₃)₂-5-CF₃CH₂]pyrimi- 5-(2,6-di-MeOPh)pyra- din-4-ylzin-2-yl 7 [2-N(CH₃)₂-5-(CH₃)₂CH]pyrimi- 2-(2,6-di-MeOPh)pyri- din-4-yldin-5-yl 8 [2-N(CH₃)₂-5-(CH₃)₂CH]pyrimi- 5-(2,6-di-MeOPh)pyri- din-4-yldin-2-yl 9 [2-N(CH₃)₂-5-(CH₃)₂CH]pyrimi- 2-(2,6-di-MeOPh)pyrida-din-4-yl zin-6-yl 10 [2-N(CH₃)₂-5-(CH₃)₂CH]pyrimi-5-(2,6-di-MeOPh)pyrimi- din-4-yl din-2-yl 11[2-N(CH₃)₂-5-(CH₃)₂CH]pyrimi- 2-(2,6-di-MeOPh)pyrimi- din-4-yl din-5-yl12 [2-N(CH₃)₂-5-(CH₃)₂CH]pyrimi- 5-(2,6-di-MeOPh)pyrazin-2-yl din-4-yl13 [2-N(CH₃)₂-5-(CH₃CH₂)₂CH]- 2-(2,6-di-MeOPh)pyridin-5-ylpyrimidin-4-yl 14 [2-N(CH₃)₂-5-(CH₃CH₂)₂CH]-5-(2,6-di-MeOPh)pyridin-2-yl pyrimidin-4-yl 15[2-N(CH₃)₂-5-(CH₃CH₂)₂CH]- 3-(2,6-di-MeOPh)pyrida- pyrimidin-4-ylzin-6-yl 16 [2-N(CH₃)₂-5-(CH₃CH₂)₂CH]- 2-(2,6-di-MeOPh)pyrimi-pyrimidin-4-yl din-2-yl 17 [2-N(CH₃)₂-5-(CH₃CH₂)₂CH]-2-(2,6-di-MeOPh)pyrimi- pyrimidin-4-yl din-5-yl 18[2-N(CH₃)₂-5-(CH₃CH₂)₂CH]- 5-(2,6-di-MeOPh)py- pyrimidin-4-ylriazin-2-yl 19 [2-N(CH₃)₂-5-(3,5-di-CH₃isoxa-2-(2,6-di-MeOPh)pyridin-5-yl zol-4-yl]pyrimidin-4-yl 20[2-N(CH₃)₂-5-(3,5-di-CH₃isoxa- 5-(2,6-di-MeOPh)pyridin-2-ylzol-4-yl]pyrimidin-4-yl 21 [2-N(CH₃)₂-5-(3,5-di-CH₃isoxa-3-(2,6-di-MeOPh)pyrida- zol-4-yl]pyrimidin-4-yl zin-6-yl 22[2-N(CH₃)₂-5-(3,5-di-CH₃isoxa- 5-(2,6-di-MeOPh)pyrimi-zol-4-yl]pyrimidin-4-yl din-2-yl 23 [2-N(CH₃)₂-5-(3,5-di-CH₃isoxa-2-(2,6-di-MeOPh)pyrimi- zol-4-yl]pyrimidin-4-yl din-5-yl 24[2-N(CH₃)₂-5-(3,5-di-CH₃isoxa- 5-(2,6-di-MeOPh)pyrazin-2-ylzol-4-yl]pyrimidin-4-yl 25 [2-N(CH₃)₂-5-(1,3,5-tri-CH₃pyra-2-(2,6-di-MeOPh)pyridin-5-yl zol-4-yl]pyrimidin-4-yl 26[2-N(CH₃)₂-5-(1,3,5-tri-CH₃pyra- 5-(2,6-di-MeOPh)pyridin-2-ylzol-4-yl]pyrimidin-4-yl 27 [2-N(CH₃)₂-5-(1,3,5-tri-CH₃pyra-3-(2,6-di-MeOPh)pyrida- zol-4-yl]pyrimidin-4-yl zin-6-yl 28[2-N(CH₃)₂-5-(1,3,5-tri-CH₃pyra- 5-(2,6-di-MeOPh)pyrimi-zol-4-yl]pyrimidin-4-yl din-2-yl 29 [2-N(CH₃)₂-5-(1,3,5-tri-CH₃pyra-2-(2,6-di-MeOPh)pyrimi- zol-4-yl]pyrimidin-4-yl din-5-yl 30[2-N(CH₃)₂-5-(1,3,5-tri-CH₃pyra- 5-(2,6-di-MeOPh)pyrazin-2-ylzol-4-yl]pyrimidin-4-yl 31 [2-N(CH₃)₂-5-(3,5-di-CH₃isothia-2-(2,6-di-MeOPh)pyridin-5-yl zol-4-yl]pyrimidin-4-yl 32[2-N(CH₃)₂-5-(3,5-di-CH₃isothia- 5-(2,6-di-MeOPh)pyridin-2-ylzol-4-yl]pyrimidin-4-yl 33 [2-N(CH₃)₂-5-(3,5-di-CH₃isothia-3-(2,6-di-MeOPh)pyrida- zol-4-yl]pyrimidin-4-yl zin-6-yl 34[2-N(CH₃)₂-5-(3,5-di-CH₃isothia- 5-(2,6-di-MeOPh)pyrimi-zol-4-yl]pyrimidin-4-yl din-2-yl 35 [2-N(CH₃)₂-5-(3,5-di-CH₃isothia-2-(2,6-di-MeOPh)pyrimi- zol-4-yl]pyrimidin-4-yl din-5-yl 36[2-N(CH₃)₂-5-(3,5-di-CH₃isothia- 5-(2,6-di-MeOPh)py-zol-4-yl]pyrimidin-4-yl riazin-2-yland are named as follows:

N-(2-(N,Ndimethylamino)-5-(2,2,2-trifluoroethyl)pyrinidin-4-yl)-L-3-(2-(2,6-dimethoxyphenyl)pyridin-5-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(2,2,2-trifluoroethyl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyridin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(2,2,2-trifluoroethyl)pyrimidin-4-yl)-L-3-(3-(2,6-dimethoxyphenyl)pyridazin-6-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(2,2,2-trifluoroethyl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyrimidin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(2,2,2-trifluoroethyl)pyrimidin-4-yl)-L-3-(2-(2,6-dimethoxyphenyl)pyridin-5-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(2-methylpropyl)pyrimidin-4-yl)-L-3-(2-(2,6-dimethoxyphenyl)pyrimidin-5-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(2,2,2-trifluoroethyl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyrazin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(2-methylpropyl)pyrimidin-4-yl)-L-3-(2-(2,6-dimethoxyphenyl)pyridin-5-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(2-methylpropyl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyridin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(2-methylpropyl)pyrimidin-4-yl)-L-3-(3-(2,6-dimethoxyphenyl)pyridazin-6-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(2-methylpropyl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyrimidin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(2-methylpropyl)pyrimidin-4-yl)-L-3-(2-(2,6-dimethoxyphenyl)pyrimidin-5-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(2-methylpropyl)pyrimidin-4-yl)-L-3-(52-(2,6-dimethoxyphenyl)pyrazin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1-ethylpropyl)pyrimidin-4-yl)-L-3-(2-(2,6-dimethoxyphenyl)pyridin-5-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1-ethylpropyl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyridin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1-ethylpropyl)pyrimidin-4-yl)-L-3-(3-(2,6-dimethoxyphenyl)pyridazin-6-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1-ethylpropyl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyrimidin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1-ethylpropyl)pyrimidin-4-yl)-L-3-(2-(2,6-dimethoxyphenyl)pyrimidin-5-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1-ethylpropyl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyrazin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(3,5-dimethylisoxazol-4-yl)pyrimidin-4-yl)-L-3-(2-(2,6-dimethoxyphenyl)pyridin-5-yl)alanine;

N-(2-(N,Ndimethylamino)-5-(3,5-dimethylisoxazol-4-yl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyridin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(3,5-dimethylisoxazol-4-yl)pyrimidin-4-yl)-L-3-(3-(2,6-dimethoxyphenyl)pyridazin-6-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(3,5-dimethylisoxazol-4-yl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyrimidin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(3,5-dimethylisoxazol-4-yl)pyrimidin-4-yl)-L-3-(2-(2,6-dimethoxyphenyl)pyrimidin-5-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(3,5-dimethylisoxazol-4-yl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyrazin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1,3,5-trimethylpyrazol-4-yl)pyrimidin-4-yl)-L-3-(2-(2,6-dimethoxyphenyl)pyridin-5-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1,3,5-trimethylpyrazol-4-yl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyridin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1,3,5-trimethylpyrazol-4-yl)pyrimidin-4-yl)-L-3-(3-(2,6-dimethoxyphenyl)pyridazin-6-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1,3,5-trimethylpyrazol-4-yl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyrimidin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1,3,5-trimethylpyrazol-4-yl)pyrimidin-4-yl)-L-3-(2-(2,6-dimethoxyphenyl)pyrimidin-5-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1,3,5-trimethylpyrazol-4-yl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyrazin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(3,5-dimethylisothiazol-4-yl)pyrimidin-4-yl)-L-3-(2-(2,6-dimethoxyphenyl)pyridin-5-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1,3,5-trimethylpyrazol-4-yl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyridin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1,3,5-trimethylpyrazol-4-yl)pyrimidin-4-yl)-L-3-(3-(2,6-dimethoxyphenyl)pyridazin-6-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1,3,5-trimethylpyrazol-4-yl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyrimidin-2-yl)alanine;

N-(2-(N,N-dimethylamino)-5-(1,3,5-trimethylpyrazol-4-yl)pyrimidin-4-yl)-L-3-(2-(2,6-dimethoxyphenyl)pyrimidin-5-yl)alanine;and

N-(2-(N,N-dimethylamino)-5-(1,3,5-trimethylpyrazol-4-yl)pyrimidin-4-yl)-L-3-(5-(2,6-dimethoxyphenyl)pyrazin-2-yl)alanine;

including all pharmaceutically acceptable salts thereof.

DETAILED DESCRIPTION OF THE INVENTION

As above, this invention relates to compounds which inhibit leukocyteadhesion and, in particular, leukocyte adhesion mediated by VLA-4.

However, prior to describing this invention in further detail, thefollowing terms will first be defined.

Definitions

As used herein, “alkyl” refers to alkyl groups preferably having from 1to 10 carbon atoms and more preferably 1 to 6 carbon atoms. This term isexemplified by groups such as methyl, t-butyl, n-heptyl, octyl and thelike.

“Alkylene” refers to alkylene groups preferably having from 1 to 4carbon atoms.

“Substituted alkyl” refers to an alkyl group, preferably of from 1 to 10carbon atoms, having from 1 to 5 substituents selected from the groupconsisting of alkoxy, substituted alkoxy, acyl, acylamino,thiocarbonylamino, acyloxy, amino, amidino, alkyl amidino, thioamidino,aminoacyl, aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy,aryl, substituted aryl, aryloxy, substituted aryloxy, aryloxyaryl,substituted aryloxyaryl, cyano, halogen, hydroxyl, nitro, carboxyl,carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl,carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-substitutedaryl, carboxylheteroaryl, carboxyl-substituted heteroaryl,carboxylheterocyclic, carboxyl-substituted heterocyclic, cycloalkyl,substituted cycloalkyl, guanidino, guanidinosulfone, thiol, thioalkyl,substituted thioalkyl, thioaryl, substituted thioaryl, thiocycloalkyl,substituted thiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,thioheterocyclic, substituted thioheterocyclic, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —OS(O)₂-alkyl, —OS(O)₂-substituted alkyl,—OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituents selectedfrom alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic and substituted heterocyclic andsubstituted alkyl groups having amino groups blocked by conventionalblocking groups such as Boc, Cbz, formyl, and the like oralkyl/substituted alkyl groups substituted with —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-alkenyl, —SO₂-substituted alkenyl,—SO₂-cycloalkyl, —SO₂-substituted cycloalkyl, —SO₂-aryl,—SO₂-substituted aryl, —SO₂-heteroaryl, —SO₂-substituted heteroaryl,—SO₂-heterocyclic, —SO₂-substituted heterocyclic and —SO₂NRR where R ishydrogen or alkyl.

“Alkoxy” refers to the group “alkyl-O—” which includes, by way ofexample, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy,sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.

“Substituted alkoxy” refers to the group “substituted alkyl-O—”.

“Alkenoxy” refers to the group “alkenyl-O—”.

“Substituted alkenoxy” refers to the group “substituted alkenyl-O—”.

“Acyl” refers to the groups H—C(O)—, alkyl-C(O)—, substitutedalkyl-C(O)—, alkenyl-C(O)—, substituted alkenyl-C(O)—, alkynyl-C(O)—,substituted alkynyl-C(O)— cycloalkyl-C(O)—, substitutedcycloalkyl-C(O)—, aryl-C(O)—, substituted aryl-C(O)—, heteroaryl-C(O)—,substituted heteroaryl-C(O), heterocyclic-C(O)—, and substitutedheterocyclic-C(O)— wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Acylamino” refers to the group —C(O)NRR where each R is independentlyselected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl,substituted aryl, cycloalkyl, substituted cycloalkyl, heteroaryl,substituted heteroaryl, heterocyclic, substituted heterocyclic and whereeach R is joined to form together with the nitrogen atom a heterocyclicor substituted heterocyclic ring wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Thiocarbonylamino” refers to the group —C(S)NRR where each R isindependently selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic and where each R is joined to form, together with thenitrogen atom a heterocyclic or substituted heterocyclic ring whereinalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic andsubstituted heterocyclic are as defined herein.

“Acyloxy” refers to the groups alkyl-C(O)O—, substituted alkyl-C(O)O—,alkenyl-C(O)O—, substituted alkenyl-C(O)O—, alkynyl-C(O)O—, substitutedalkynyl-C(O)O—, aryl-C(O)O—, substituted aryl-C(O)O—, cycloalkyl-C(O)O—,substituted cycloalkyl-C(O)O—, heteroaryl-C(O)O—, substitutedheteroaryl-C(O)O—, heterocyclic-C(O)O—, and substitutedheterocyclic-C(O)O— wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Oxysulfonyl” refers to the groups alkyl-SO₂O—, substituted alkyl-SO₂O—,alkenyl-SO₂O—, substituted alkenyl-SO₂O—, alkynyl-SO₂O—, substitutedalkynyl-SO₂O—, aryl-SO₂O—, substituted aryl-SO₂O—, cycloalkyl-SO₂O—,substituted cycloalkyl-SO₂O—, heteroaryl-SO₂O—, substitutedheteroaryl-SO₂O—, heterocyclic-SO₂O—, and substituted heterocyclic-SO₂O—wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic andsubstituted heterocyclic are as defined herein.

“Alkenyl” refers to alkenyl group preferably having from 2 to 10 carbonatoms and more preferably 2 to 6 carbon atoms and having at least 1 andpreferably from 1-2 sites of alkenyl unsaturation.

“Substituted alkenyl” refers to alkenyl groups having from 1 to 5substituents selected from the group consisting of alkoxy, substitutedalkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, amino, amidino,alkylamidino, thioamidino, aminoacyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aryl, substituted aryl,aryloxy, substituted aryloxy, aryloxyaryl, substituted aryloxyaryl,halogen, hydroxyl, cyano, nitro, carboxyl, carboxylalkyl,carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-substitutedcycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl,carboxyl-substituted heteroaryl, carboxylheterocyclic,carboxyl-substituted heterocyclic, cycloalkyl, substituted cycloalkyl,guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl,thioaryl, substituted thioaryl, thiocycloalkyl, substitutedthiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,thioheterocyclic, substituted thioheterocyclic, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —OS(O)₂-alkyl, —OS(O)₂-substituted alkyl,—OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituents selectedfrom alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic and substituted heterocyclic andsubstituted alkenyl groups having amino groups blocked by conventionalblocking groups such as Boc, Cbz, formyl, and the like oralkenyl/substituted alkenyl groups substituted with —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-alkenyl, —SO₂-substituted alkenyl,—SO₂-cycloalkyl, —SO₂-substituted cycloalkyl, —SO₂-aryl,—SO₂-substituted aryl, —SO₂-heteroaryl, —SO₂-substituted heteroaryl,—SO₂-heterocyclic, —SO₂-substituted heterocyclic and —SO₂NRR where R ishydrogen or alkyl.

“Alkynyl” refers to alkynyl group preferably having from 2 to 10 carbonatoms and more preferably 3 to 6 carbon atoms and having at least 1 andpreferably from 1-2 sites of alkynyl unsaturation.

“Substituted alkynyl” refers to alkynyl groups having from 1 to 5substituents selected from the group consisting of alkoxy, substitutedalkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, amino, amidino,alkylamidino, thioamidino, aminoacyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aryl, substituted aryl,aryloxy, substituted aryloxy, aryloxyaryl, substituted aryloxyaryl,halogen, hydroxyl, cyano, nitro, carboxyl, carboxylalkyl,carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-substitutedcycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl,carboxyl-substituted heteroaryl, carboxylheterocyclic,carboxyl-substituted heterocyclic, cycloalkyl, substituted cycloalkyl,guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl,thioaryl, substituted thioaryl, thiocycloalkyl, substitutedthiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,thioheterocyclic, substituted thioheterocyclic, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —OS(O)₂-alkyl, —OS(O)₂-substituted alkyl,—OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituents selectedfrom alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic and substituted heterocyclic andsubstituted alkynyl groups having amino groups blocked by conventionalblocking groups such as Boc, Cbz, formyl, and the like oralkynyl/substituted alkynyl groups substituted with —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-alkenyl, —SO₂-substituted alkenyl,—SO₂cycloalkyl, —SO₂-substituted cycloalkyl, —SO₂-aryl, —SO₂-substitutedaryl, —SO₂-heteroaryl, —SO₂-substituted heteroaryl, —SO₂-heterocyclic,—SO₂-substituted heterocyclic and —SO₂NRR where R is hydrogen or alkyl.

“Amidino” refers to the group H₂NC(═NH)— and the term “alkylamidino”refers to compounds having 1 to 3 alkyl groups (e.g., alkylHNC(═NH)—).

“Thioamidino” refers to the group RSC(═NH)— where R is hydrogen oralkyl.

“Amino” refers to the group —NH₂.

“Substituted amino” refers to the group —NRR, where each R group isindependently selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-alkenyl,—SO₂-substituted alkenyl, —SO₂-cycloalkyl, —SO₂-substituted cycloalkyl,—SO₂-aryl, —SO₂-substituted aryl, —SO₂-heteroaryl, —SO₂-substitutedheteroaryl, —SO₂-heterocyclic, —SO₂-substituted heterocyclic, providedthat both R groups are not hydrogen; or the R groups can be joinedtogether with the nitrogen atom to form a heterocyclic or substitutedheterocyclic ring.

“Aminoacyl” refers to the groups —NRC(O)alkyl, —NRC(O)substituted alkyl,—NRC(O)cycloalkyl, —NRC(O)substituted cycloalkyl, —NRC(O)alkenyl,—NRC(O)substituted alkenyl, —NRC(O)alkynyl, —NRC(O)substituted alkynyl,—NRC(O)aryl, —NRC(O)substituted aryl, —NRC(O)heteroaryl,—NRC(O)substituted heteroaryl, —NRC(O)heterocyclic, and—NRC(O)substituted heterocyclic where R is hydrogen or alkyl and whereinalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic andsubstituted heterocyclic are as defined herein.

“Aminosulfonyl” refers to the groups —NRSO₂alkyl, —NRSO₂substitutedalkyl, —NRSO₂cycloalkyl, —NRSO₂substituted cycloalkyl, —NRSO₂alkenyl,—NRSO₂substituted alkenyl, —NRSO₂alkynyl, —NRSO₂substituted alkynyl,—NRSO₂aryl, —NRSO₂substituted aryl, —NRSO₂heteroaryl, —NRSO₂substitutedheteroaryl, —NRSO₂heterocyclic, and —NRSO₂substituted heterocyclic whereR is hydrogen or alkyl and wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Aminocarbonyloxy” refers to the groups —NRC(O)O-alkyl,—NRC(O)O-substituted alkyl, —NRC(O)O-alkenyl, —NRC(O)O-substitutedalkenyl, —NRC(O)O-alkynyl, —NRC(O)O-substituted alkynyl,—NRC(O)O-cycloalkyl, —NRC(O)O-substituted cycloalkyl, —NRC(O)O-aryl,—NRC(O)O-substituted aryl, —NRC(O)O-heteroaryl, —NRC(O)O-substitutedheteroaryl, —NRC(O)O-heterocyclic, and —NRC(O)O-substituted heterocyclicwhere R is hydrogen or alkyl and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Aminosulfonyloxy” refers to the groups —NRSO₂O-alkyl,—NRSO₂O-substituted alkyl, —NRSO₂O-alkenyl, —NRSO₂O-substituted alkenyl,—NRSO₂O-alkynyl, —NRSO₂O-substituted alkynyl, —NRSO₂O-cycloalkyl,—NRSO₂O-substituted cycloalkyl, —NRSO₂O-aryl, —NRSO₂O-substituted aryl,—NRSO₂O-heteroaryl, —NRSO₂O-substituted heteroaryl,—NRSO₂O-heterocyclic, and —NRSO₂O-substituted heterocyclic where R ishydrogen or alkyl and wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Oxycarbonylamino” refers to the groups —OC(O)NH₂, —OC(O)NRR,—OC(O)NR-alkyl, —OC(O)NR-substituted alkyl, —OC(O)NR-alkenyl,—OC(O)NR-substituted alkenyl, —OC(O)NR-alkynyl, —OC(O)NR-substitutedalkynyl, —OC(O)NR-cycloalkyl, —OC(O)NR-substituted cycloalkyl,—OC(O)NR-aryl, —OC(O)NR-substituted aryl, —OC(O)NR-heteroaryl,—OC(O)NR-substituted heteroaryl, —OC(O)NR-heterocyclic, and—OC(O)NR-substituted heterocyclic where R is hydrogen, alkyl or whereeach R is joined to form, together with the nitrogen atom a heterocyclicor substituted heterocyclic ring and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Oxythiocarbonylamino” refers to the groups —OC(S)NH₂, —OC(S)NRR,—OC(S)NR-alkyl, —OC(S)NR-substituted alkyl, —OC(S)NR-alkenyl,—OC(S)NR-substituted alkenyl, —OC(S)NR-alkynyl, —OC(S)NR-substitutedalkynyl, —OC(S)NR-cycloalkyl, —OC(S)NR-substituted cycloalkyl,—OC(S)NR-aryl, —OC(S)NR-substituted aryl, —OC(S)NR-heteroaryl,—OC(S)NR-substituted heteroaryl, —OC(S)NR-heterocyclic, and—OC(S)NR-substituted heterocyclic where R is hydrogen, alkyl or whereeach R is joined to form together with the nitrogen atom a heterocyclicor substituted heterocyclic ring and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Oxysulfonylamino” refers to the groups —OSO₂NH₂, —OSO₂NRR,—OSO₂NR-alkyl, —OSO₂NR-substituted alkyl, —OSO₂NR-alkenyl,—OSO₂NR-substituted alkenyl, —OSO₂NR-alkynyl, —OSO₂NR-substitutedalkynyl, —OSO₂NR-cycloalkyl, —OSO₂NR-substituted cycloalkyl,—OSO₂NR-aryl, —OSO₂NR-substituted aryl, —OSO₂NR-heteroaryl,—OSO₂NR-substituted heteroaryl, —OSO₂NR-heterocyclic, and—OSO₂NR-substituted heterocyclic where R is hydrogen, alkyl or whereeach R is joined to form, together with the nitrogen atom a heterocyclicor substituted heterocyclic ring and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Aminocarbonylamino” refers to the groups —NRC(O)NRR, —NRC(O)NR-alkyl,—NRC(O)NR-substituted alkyl, —NRC(O)NR-alkenyl, —NRC(O)NR-substitutedalkenyl, —NRC(O)NR-alkynyl, —NRC(O)NR-substituted alkynyl,—NRC(O)NR-aryl, —NRC(O)NR-substituted aryl, —NRC(O)NR-cycloalkyl,—NRC(O)NR-substituted cycloalkyl, —NRC(O)NR-heteroaryl, and—NRC(O)NR-substituted heteroaryl, —NRC(O)NR-heterocyclic, and—NRC(O)NR-substituted heterocyclic where each R is independentlyhydrogen, alkyl or where each R is joined to form together with thenitrogen atom a heterocyclic or substituted heterocyclic ring as well aswhere one of the amino groups is blocked by conventional blocking groupssuch as Boc, Cbz, formyl, and the like and wherein alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic and substituted heterocyclic are asdefined herein.

“Aminothiocarbonylamino” refers to the groups —NRC(S)NRR,—NRC(S)NR-alkyl, —NRC(S)NR-substituted alkyl, —NRC(S)NR-alkenyl,—NRC(S)NR-substituted alkenyl, —NRC(S)NR-alkynyl, —NRC(S)NR-substitutedalkynyl, —NRC(S)NR-aryl, —NRC(S)NR-substituted aryl,—NRC(S)NR-cycloalkyl, —NRC(S)NR-substituted cycloalkyl,—NRC(S)NR-heteroaryl, and —NRC(S)NR-substituted heteroaryl,—NRC(S)NR-heterocyclic, and —NRC(S)NR-substituted heterocyclic whereeach R is independently hydrogen, alkyl or where each R is joined toform together with the nitrogen atom a heterocyclic or substitutedheterocyclic ring as well as where one of the amino groups is blocked byconventional blocking groups such as Boc, Cbz, formyl, and the like andwherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic andsubstituted heterocyclic are as defined herein.

“Aminosulfonylamino” refers to the groups —NRSO₂NRR, —NRSO₂NR-alkyl,—NRSO₂NR-substituted alkyl, —NRSO₂NR-alkenyl, —NRSO₂NR-substitutedalkenyl, —NRSO₂NR-alkynyl, —NRSO₂NR-substituted alkynyl, —NRSO₂NR-aryl,—NRSO₂NR-substituted aryl, —NRSO₂NR-cycloalkyl, —NRSO₂NR-substitutedcycloalkyl, —NRSO₂NR-heteroaryl, and —NRSO₂NR-substituted heteroaryl,—NRSO₂NR-heterocyclic, and —NRSO₂NR-substituted heterocyclic, where eachR is independently hydrogen, alkyl or where each R is joined to formtogether with the nitrogen atom a heterocyclic or substitutedheterocyclic ring as well as where one of the amino groups is blocked byconventional blocking groups such as Boc, Cbz, formyl, and the like andwherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic andsubstituted heterocyclic are as defined herein.

“Aryl” or “Ar” refers to an unsaturated aromatic carbocyclic group offrom 6 to 14 carbon atoms having a single ring (e.g., phenyl) ormultiple condensed rings (e.g., naphthyl or anthryl) which condensedrings may or may not be aromatic (e.g., 2-benzoxazolinone,2H-1,4-benzoxazin-3(4H)-one-7yl, and the like). Preferred aryls includephenyl and naphthyl.

Substituted aryl refers to aryl groups which are substituted with from 1to 3 substituents selected from the group consisting of hydroxy, acyl,acylamino, thiocarbonylamino, acyloxy, alkyl, substituted alkyl, alkoxy,substituted alkoxy, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, amidino, alkylamidino, thioamidino, amino, aminoacyl,aminocarbonyloxy, aminocarbonylamino, aminothiocarbonylamino, aryl,substituted aryl, aryloxy, substituted aryloxy, cycloalkoxy, substitutedcycloalkoxy, heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy,substituted heterocyclyloxy, carboxyl, carboxylalkyl,carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-substitutedcycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl,carboxyl-substituted heteroaryl, carboxylheterocyclic,carboxyl-substituted heterocyclic, carboxylamido, cyano, thiol,thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl,thioheteroaryl, substituted thioheteroaryl, thiocycloalkyl, substitutedthiocycloalkyl, thioheterocyclic, substituted thioheterocyclic,cycloalkyl, substituted cycloalkyl, guanidino, guanidinosulfone, halo,nitro, heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy,substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,oxycarbonylamino, oxythiocarbonylamino, —S(O)₂-alkyl, —S(O)₂-substitutedalkyl, —S(O)₂-cycloalkyl, —S(O)₂-substituted cycloalkyl, —S(O)₂-alkenyl,—S(O)₂-substituted alkenyl, —S(O)₂-aryl, —S(O)₂-substituted aryl,—S(O)₂-heteroaryl, —S(O)₂-substituted heteroaryl, —S(O)₂-heterocyclic,—S(O)₂-substituted heterocyclic, —OS(O)₂-alkyl, —OS(O)₂-substitutedalkyl, —OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituents selectedfrom alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic and substituted heterocyclic andamino groups on the substituted aryl blocked by conventional blockinggroups such as Boc, Cbz, formyl, and the like or substituted with—SO₂NRR where R is hydrogen or alkyl.

“Aryloxy” refers to the group aryl-O— which includes, by way of example,phenoxy, naphthoxy, and the like.

“Substituted aryloxy” refers to substituted aryl-O— groups.

“Aryloxyaryl” refers to the group -aryl-O-aryl.

“Substituted aryloxyaryl” refers to aryloxyaryl groups substituted withfrom 1 to 3 substituents on either or both aryl rings selected from thegroup consisting of hydroxy, acyl, acylamino, thiocarbonylamino,acyloxy, alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, amidino,alkylamidino, thioamidino, amino, aminoacyl, aminocarbonyloxy,aminocarbonylamino, aminothiocarbonylamino, aryl, substituted aryl,aryloxy, substituted aryloxy, cycloalkoxy, substituted cycloalkoxy,heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substitutedheterocyclyloxy, carboxyl, carboxylalkyl, carboxyl-substituted alkyl,carboxyl-cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl,carboxyl-substituted aryl, carboxylheteroaryl, carboxyl-substitutedheteroaryl, carboxylheterocyclic, carboxyl-substituted heterocyclic,carboxylamido, cyano, thiol, thioalkyl, substituted thioalkyl, thioaryl,substituted thioaryl, thioheteroaryl, substituted thioheteroaryl,thiocycloalkyl, substituted thiocycloalkyl, thioheterocyclic,substituted thioheterocyclic, cycloalkyl, substituted cycloalkyl,guanidino, guanidinosulfone, halo, nitro, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —S(O)₂-alkyl, —S(O)₂-substituted alkyl,—S(O)₂-cycloalkyl, —S(O)₂-substituted cycloalkyl, —S(O)₂-alkenyl,—S(O)₂-substituted alkenyl, —S(O)₂-aryl, —S(O)₂-substituted aryl,—S(O)₂-heteroaryl, —S(O)₂-substituted heteroaryl, —S(O)₂-heterocyclic,—S(O)₂-substituted heterocyclic, —OS(O)₂-alkyl, —OS(O)₂-substitutedalkyl, —OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituents selectedfrom alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic and substituted heterocyclic andamino groups on the substituted aryl blocked by conventional blockinggroups such as Boc, Cbz, formyl, and the like or substituted with—SO₂NRR where R is hydrogen or alkyl.

“Cycloalkyl” refers to cyclic alkyl groups of from 3 to 8 carbon atomshaving a single cyclic ring including, by way of example, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cyclooctyl and the like. Excludedfrom this definition are multi-ring alkyl groups such as adamantanyl,etc.

“Cycloalkenyl” refers to cyclic alkenyl groups of from 3 to 8 carbonatoms having single or multiple unsaturation but which are not aromatic.

“Substituted cycloalkyl” and “substituted cycloalkenyl” refer to acycloalkyl and cycloalkenyl groups, preferably of from 3 to 8 carbonatoms, having from 1 to 5 substituents selected from the groupconsisting of oxo (═O), thioxo (═S), alkoxy, substituted alkoxy, acyl,acylamino, thiocarbonylamino, acyloxy, amino, amidino, alkylamidino,thioamidino, aminoacyl, aminocarbonylamino, aminothiocarbonylamino,aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy,aryloxyaryl, substituted aryloxyaryl, halogen, hydroxyl, cyano, nitro,carboxyl, carboxylalkyl, carboxyl-substituted alkyl,carboxyl-cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl,carboxyl-substituted aryl, carboxylheteroaryl, carboxyl-substitutedheteroaryl, carboxylheterocyclic, carboxyl-substituted heterocyclic,cycloalkyl, substituted cycloalkyl, guanidino, guanidinosulfone, thiol,thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl,thiocycloalkyl, substituted thiocycloalkyl, thioheteroaryl, substitutedthioheteroaryl, thioheterocyclic, substituted thioheterocyclic,heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy,substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,oxycarbonylamino, oxythiocarbonylamino, —OS(O)₂-alkyl,—OS(O)₂-substituted alkyl, —OS(O)₂-aryl, —OS(O)₂-substituted aryl,—OS(O)₂-heteroaryl, —OS(O)₂-substituted heteroaryl,—OS(O)₂-heterocyclic, —OS(O)₂-substituted heterocyclic, —OSO₂—NRR whereR is hydrogen or alkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl,—NRS(O)₂-aryl, —NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl,—NRS(O)₂-substituted heteroaryl, —NRS(O)₂—heterocyclic,—NRS(O)₂-substituted heterocyclic, —NRS(O)₂—NR-alkyl,—NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl, —NRS(O)₂—NR-substitutedaryl, —NRS(O)₂—NR-heteroaryl, —NRS(O)₂—NR-substituted heteroaryl,—NRS(O)₂—NR-heterocyclic, —NRS(O)₂—NR-substituted heterocyclic where Ris hydrogen or alkyl, mono- and di-alkylamino, mono- and di-(substitutedalkyl)amino, mono- and di-arylamino, mono- and di-substituted arylamino,mono- and di-heteroarylamino, mono- and di-substituted heteroarylamino,mono- and di-heterocyclic amino, mono- and di-substituted heterocyclicamino, unsymmetric di-substituted amines having different substituentsselected from alkyl, substituted alkyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic and substitutedheterocyclic and substituted alkynyl groups having amino groups blockedby conventional blocking groups such as Boc, Cbz, formyl, and the likeor alkynyl/substituted alkynyl groups substituted with —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-alkenyl, —SO₂-substituted alkenyl,—SO₂-cycloalkyl, —SO₂-substituted cycloalkyl, —SO₂-aryl,—SO₂-substituted aryl, —SO₂-heteroaryl, —SO₂-substituted heteroaryl,—SO₂-heterocyclic, —SO₂-substituted heterocyclic and —SO₂NRR where R ishydrogen or alkyl.

“Cycloalkoxy” refers to —O-cycloalkyl groups.

“Substituted cycloalkoxy” refers to —O-substituted cycloalkyl groups.

“Cycloalkenoxy” refers to —O-cycloalkenyl groups.

“Substituted cycloalkenoxy” refers to —O-substituted cycloalkenylgroups.

“Guanidino” refers to the groups —NRC(═NR)NRR, —NRC(═NR)NR-alkyl,—NRC(═NR)NR-substituted alkyl, —NRC(═NR)NR-alkenyl,—NRC(═NR)NR-substituted alkenyl, —NRC(═NR)NR-alkynyl,—NRC(═NR)NR-substituted alkynyl, —NRC(═NR)NR-aryl,—NRC(═NR)NR-substituted aryl, —NRC(═NR)NR-cycloalkyl,—NRC(═NR)NR-heteroaryl, —NRC(═NR)NR-substituted heteroaryl,—NRC(═NR)NR-heterocyclic, and —NRC(═NR)NR-substituted heterocyclic whereeach R is independently hydrogen and alkyl as well as where one of theamino groups is blocked by conventional blocking groups such as Boc,Cbz, formyl, and the like and wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Guanidinosulfone” refers to the groups —NRC(═NR)NRSO₂-alkyl,—NRC(═NR)NRSO₂-substituted alkyl, —NRC(═NR)NRSO₂-alkenyl,—NRC(═NR)NRSO₂-substituted alkenyl, —NRC(═NR)NRSO₂-alkynyl,—NRC(═NR)NRSO₂-substituted alkynyl, —NRC(═NR)NRSO₂-aryl,—NRC(═NR)NRSO₂-substituted aryl, —NRC(═NR)NRSO₂-cycloalkyl,—NRC(═NR)NRSO₂-substituted cycloalkyl, —NRC(═NR)NRSO₂-heteroaryl, and—NRC(═NR)NRSO₂-substituted heteroaryl, —NRC(═NR)NRSO₂-heterocyclic, and—NRC(═NR)NRSO₂-substituted heterocyclic where each R is independentlyhydrogen and alkyl and wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Halo” or “halogen” refers to fluoro, chloro, bromo and iodo andpreferably is either chloro or bromo.

“Heteroaryl” refers to an aromatic carbocyclic group of from 2 to 10carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen andsulfur within the ring or oxides thereof. Such heteroaryl groups canhave a single ring (e.g., pyridyl or furyl) or multiple condensed rings(e.g., indolizinyl or benzothienyl). Additionally, the heteroatoms ofthe heteroaryl group may be oxidized, i.e., to form pyridine N-oxides or1,1-dioxo-1,2,5-thiadiazoles and the like. Preferred heteroaryls includepyridyl, pyrrolyl, indolyl, furyl, pyridazinyl, pyrimidinyl, pyrazinyl,1-oxo-1,2,5-thiadiazolyl and 1,1-dioxo-1,2,5-thiadiazolyl. The term“heteroaryl having two nitrogen atoms in the heteroaryl ring” refers toa heteroaryl group having two, and only two, nitrogen atoms in theheteroaryl ring and optionally containing 1 or 2 other heteroatoms inthe heteroaryl ring, such as oxygen or sulfur

“Substituted heteroaryl” refers to heteroaryl groups which aresubstituted with from 1 to 3 substituents selected from the groupconsisting of hydroxy, acyl, acylamino, thiocarbonylamino, acyloxy,alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, amidino,alkylamidino, thioamidino, amino, aminoacyl, aminocarbonyloxy,aminocarbonylamino, aminothiocarbonylamino, aryl, substituted aryl,aryloxy, substituted aryloxy, cycloalkoxy, substituted cycloalkoxy,heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substitutedheterocyclyloxy, carboxyl, carboxylalkyl, carboxyl-substituted alkyl,carboxyl-cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl,carboxyl-substituted aryl, carboxylheteroaryl, carboxyl-substitutedheteroaryl, carboxylheterocyclic, carboxyl-substituted heterocyclic,carboxylamido, cyano, thiol, thioalkyl, substituted thioalkyl, thioaryl,substituted thioaryl, thioheteroaryl, substituted thioheteroaryl,thiocycloalkyl, substituted thiocycloalkyl, thioheterocyclic,substituted thioheterocyclic, cycloalkyl, substituted cycloalkyl,guanidino, guanidinosulfone, halo, nitro, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —S(O)₂-alkyl, —S(O)₂-substituted alkyl,—S(O)₂-cycloalkyl, —S(O)₂-substituted cycloalkyl, —S(O)₂-alkenyl,—S(O)₂-substituted alkenyl, —S(O)₂-aryl, —S(O)₂-substituted aryl,—S(O)₂-heteroaryl, —S(O)₂-substituted heteroaryl, —S(O)₂-heterocyclic,—S(O)₂-substituted heterocyclic, —OS(O)₂-alkyl, —OS(O)₂-substitutedalkyl, —OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituents selectedfrom alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic and substituted heterocyclic andamino groups on the substituted aryl blocked by conventional blockinggroups such as Boc, Cbz, formyl, and the like or substituted with—SO₂NRR where R is hydrogen or alkyl.

“Nitrogen containing heteroaryl” refers to a heteroary ring as definedabove that contains at least one nitrogen atom in the ring. Suchheteroaryl groups can have a single ring (e.g., pyridyl) or multiplecondensed rings (e.g., indolizinyl). Additionally, the heteroatoms ofthe heteroaryl group may be oxidized, i.e., to form pyridine N-oxides or1,1-dioxo-1,2,5-thiadiazoles and the like. Preferred nitrogen containingheteroaryls include pyridyl, pyrrolyl, indolyl, pyridazinyl,pyrimidinyl, pyrazinyl, 1-oxo-1,2,5-thiadiazolyl and 1,1-dioxo-1,2,5-thiadiazolyl. The term “heteroaryl having two nitrogenatoms in the heteroaryl ring” refers to a heteroaryl group having two,and only two, nitrogen atoms in the heteroaryl ring and optionallycontaining 1 or 2 other heteroatoms in the heteroaryl ring, such asoxygen or sulfur

“Nitrogen containing substituted heteroaryl” refers to heteroaryl groupsas defined above which are substituted with from 1 to 3 substituentsselected from the group consisting of hydroxy, acyl, acylamino,thiocarbonylamino, acyloxy, alkyl, substituted alkyl, alkoxy,substituted alkoxy, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, amidino, alkylamidino, thioamidino, amino, aminoacyl,aminocarbonyloxy, aminocarbonylamino, aminothiocarbonylamino, aryl,substituted aryl, aryloxy, substituted aryloxy, cycloalkoxy, substitutedcycloalkoxy, heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy,substituted heterocyclyloxy, carboxyl, carboxylalkyl,carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-substitutedcycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl,carboxyl-substituted heteroaryl, carboxytheterocyclic,carboxyl-substituted heterocyclic, carboxylamido, cyano, thiol,thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl,thioheteroaryl, substituted thioheteroaryl, thiocycloalkyl, substitutedthiocycloalkyl, thioheterocyclic, substituted thioheterocyclic,cycloalkyl, substituted cycloalkyl, guanidino, guanidinosulfone, halo,nitro, heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy,substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,oxycarbonylamino, oxythiocarbonylamino, —S(O)₂-alkyl, —S(O)₂-substitutedalkyl, —S(O)₂-cycloalkyl, —S(O)₂-substituted cycloalkyl, —S(O)₂-alkenyl,—S(O)₂-substituted alkenyl, —S(O)₂-aryl, —S(O)₂-substituted aryl,—S(O)₂-heteroaryl, —S(O)₂-substituted heteroaryl, —S(O)₂-heterocyclic,—S(O)₂-substituted heterocyclic, —OS(O)₂-alkyl, —OS(O)₂-substitutedalkyl, —OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituents selectedfrom alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic and substituted heterocyclic andamino groups on the substituted aryl blocked by conventional blockinggroups such as Boc, Cbz, formyl, and the like or substituted with—SO₂NRR where R is hydrogen or alkyl.

“Heteroaryloxy” refers to the group —O-heteroaryl and “substitutedheteroaryloxy” refers to the group —O-substituted heteroaryl.

“Heterocycle” or “heterocyclic” refers to a saturated or unsaturatedgroup having a single ring or multiple condensed rings, from 1 to 10carbon atoms and from 1 to 4 hetero atoms selected from nitrogen, sulfuror oxygen within the ring wherein, in fused ring systems, one or more ofthe rings can be aryl or heteroaryl.

“Substituted heterocyclic” refers to heterocycle groups which aresubstituted with from 1 to 3 substituents selected from the groupconsisting of oxo (═O), thioxo (═S), alkoxy, substituted alkoxy, acyl,acylamino, thiocarbonylamino, acyloxy, amino, amidino, alkylamidino,thioamidino, aminoacyl, aminocarbonylamino, aminothiocarbonylamino,aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy,aryloxyaryl, substituted aryloxyaryl, halogen, hydroxyl, cyano, nitro,carboxyl, carboxylalkyl, carboxyl-substituted alkyl,carboxyl-cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl,carboxyl-substituted aryl, carboxylheteroaryl, carboxyl-substitutedheteroaryl, carboxylheterocyclic, carboxyl-substituted heterocyclic,cycloalkyl, substituted cycloalkyl, guanidino, guanidinosulfone, thiol,thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl,thiocycloalkyl, substituted thiocycloalkyl, thioheteroaryl, substitutedthioheteroaryl, thioheterocyclic, substituted thioheterocyclic,heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy,substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,oxycarbonylamino, oxythiocarbonylamino, —OS(O)₂-alkyl,—OS(O)₂-substituted alkyl, —OS(O)₂-aryl, —OS(O)₂-substituted aryl,—OS(O)₂-heteroaryl, —OS(O)₂-substituted heteroaryl,—OS(O)₂-heterocyclic, —OS(O)₂-substituted heterocyclic, —OSO₂—NRR whereR is hydrogen or alkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl,—NRS(O)₂-aryl, —NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl,—NRS(O)₂-substituted heteroaryl, —NRS(O)₂-heterocyclic,—NRS(O)₂-substituted heterocyclic, —NRS(O)₂—NR-alkyl,—NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl, —NRS(O)₂—NR-substitutedaryl, —NRS(O)₂—NR-heteroaryl, —NRS(O)₂—NR-substituted heteroaryl,—NRS(O)₂—NR-heterocyclic, —NRS(O)₂—NR-substituted heterocyclic where Ris hydrogen or alkyl, mono- and di-alkylamino, mono- and di-(substitutedalkyl)amino, mono- and di-arylamino, mono- and di-substituted arylamino,mono- and di-heteroarylamino, mono- and di-substituted heteroarylamino,mono- and di-heterocyclic amino, mono- and di-substituted heterocyclicamino, unsymmetric di-substituted amines having different substituentsselected from alkyl, substituted alkyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic and substitutedheterocyclic and substituted alkynyl groups having amino groups blockedby conventional blocking groups such as Boc, Cbz, formyl, and the likeor alkynyl/substituted alkynyl groups substituted with —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-alkenyl, —SO₂-substituted alkenyl,—SO₂-cycloalkyl, —SO₂-substituted cycloalkyl, —SO₂-aryl,—SO₂-substituted aryl, —SO₂-heteroaryl, —SO₂-substituted heteroaryl,—SO₂-heterocyclic, —SO₂-substituted heterocyclic and —SO₂NRR where R ishydrogen or alkyl.

Examples of heterocycles and heteroaryls include, but are not limitedto, azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine,pyrimidine, pyridazine, indolizine, isoindole, indole, dihydroindole,indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine,naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine,carbazole, carboline, phenanthridine, acridine, phenanthroline,isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine,imidazolidine, imidazoline, piperidine, piperazine, indoline,phthalimide, 1,2,3,4-tetrahydroisoquinoline,4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene,benzo[b]thiophene, morpholino, thiomorpholino, piperidinyl, pyrrolidine,tetrahydrofuranyl, and the like.

“Heterocyclyloxy” refers to the group —O-heterocyclic and “substitutedheterocyclyloxy” refers to the group —O-substituted heterocyclic.

“Thiol” refers to the group —SH.

“Thioalkyl” refers to the groups —S-alkyl

“Substituted thioalkyl” refers to the group —S-substituted alkyl.

“Thiocycloalkyl” refers to the groups —S-cycloalkyl.

“Substituted thiocycloalkyl” refers to the group —S-substitutedcycloalkyl.

“Thioaryl” refers to the group —S-aryl and “substituted thioaryl” refersto the group —S-substituted aryl.

“Thioheteroaryl” refers to the group —S-heteroaryl and “substitutedthioheteroaryl” refers to the group —S-substituted heteroaryl.

“Thioheterocyclic” refers to the group —S-heterocyclic and “substitutedthioheterocyclic” refers to the group —S-substituted heterocyclic.

“Pharmaceutically acceptable salt” refers to pharmaceutically acceptablesalts of a compound of Formula (I) which salts are derived from avariety of organic and inorganic counter ions well known in the art andinclude, by way of example only, sodium, potassium, calcium, magnesium,ammonium, tetraalkylammonium, and the like; and when the moleculecontains a basic functionality, salts of organic or inorganic acids,such as hydrochloride, hydrobromide, tartrate, mesylate, acetate,maleate, oxalate and the like.

General Synthetic Scheme

The compounds of this invention can be prepared from readily availablestarting materials using the following general methods and procedures.It will be appreciated that where typical or preferred processconditions (i.e., reaction temperatures, times, mole ratios ofreactants, solvents, pressures, etc.) are given, other processconditions can also be used unless otherwise stated. Optimum reactionconditions may vary with the particular reactants or solvent used, butsuch conditions can be determined by one skilled in the art by routineoptimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. Suitableprotecting groups for various functional groups as well as suitableconditions for protecting and deprotecting particular functional groupsare well known in the art. For example, numerous protecting groups aredescribed in T. W. Greene and G. M. Wuts, Protecting Groups in OrganicSynthesis, Second Edition, Wiley, N.Y., 1991, and references citedtherein.

Furthermore, the compounds of this invention will typically contain oneor more chiral centers. Accordingly, if desired, such compounds can beprepared or isolated as pure stereoisomers, i.e., as individualenantiomers or diastereomers, or as stereoisomer-enriched mixtures. Allsuch stereoisomers (and enriched mixtures) are included within the scopeof this invention, unless otherwise indicated. Pure stereoisomers (orenriched mixtures) may be prepared using, for example, optically activestarting materials or stereoselective reagents well-known in the art.Alternatively, racemic mixtures of such compounds can be separatedusing, for example, chiral column chromatography, chiral resolvingagents and the like.

In a preferred method of synthesis, the compounds of this invention areprepared by coupling an amino acid derivative of the formula (a):

where HetAr is as defined herein and P¹ is a carboxylic acid protectinggroup (such as an alkyl group, i.e. methyl, ethyl and the like), with asuitably functionalized cyclic group (e.g., heteroaryl or heterocyclicintermediate). For example, such coupling reactions may be performed bydisplacing a leaving group, such as chloro, bromo, iodo, tosyl and thelike, from the cyclic intermediate, e.g., heteroaryl or heterocyclicintermediate, with the amino group of the amino acid derivative; or byreductive alkylation of the amino group of amino acid derivative with acarbonyl-functionalized intermediate. Such coupling reactions arewell-known to those skilled in the art.

By way of illustration, the synthesis of a representative compound ofFormula (I) is shown in Scheme 1.

As shown in Scheme 1, 5-nitrouracil, 1, (commercially available fromAldrich Chemical Company, Milwaukee, Wis. USA) is treated withphosphorus oxychloride and N,N-dimethylaniline according to theprocedure described in Whittaker, J. Chem. Soc. 1951, 1565 to give1,3-dichloro-4-nitropyrimidine, 2.

1,3-Dichloro-4-nitropyrimidine, 2, is then reacted with about one molarequivalent of an amino acid derivative of the formula:H₂N—CH(CH₂HetAr)C(O)X where HetAr and X are as defined herein or X is—OP¹ where P¹ is a carboxylic acid protecting group, in the presence ofa trialkcylamine, such as diisopropylethylamine (DIEA). Typically, thisreaction is conducted in an inert diluent, such as dichloromethane, at atemperature ranging from about 0° C. to about 10° C. for about 5 min. toabout 6 hours to afford intermediate 3.

The nitro group of intermediate 3 is then reduced using a conventionalreducing agent, such as hydrogen and a palladium on carbon catalyst.When hydrogen and palladium on carbon are employed as the reducingagent, the chloro group of intermediate 3 is also removed. This reactionis typically conducted by contacting 3 with a Degussa-type palladium oncarbon catalyst (typically 20%) and excess sodium bicarbonate in aninert diluent, such as methanol, under hydrogen (typically about 55 psi)for about 12 to 36 hours at ambient temperature to afford aminointermediate 4.

Amino intermediate 4 is then reacted with a sulfonyl chloride of theformula: R⁵—S(O)₂—Cl, where R⁵ is as defined herein, to providesulfonamide intermediate 5. This reaction is typically conducted byreacting the amino intermediate 4 with at least one equivalent,preferably about 1.1 to about 2 equivalents, of the sulfonyl chloride inan inert diluent such as dichloromethane and the like. Generally, thereaction is conducted at a temperature ranging from about −70° C. toabout 40° C. for about 1 to about 24 hours. Preferably, this reaction isconducted in the presence of a suitable base to scavenge the acidgenerated during the reaction. Suitable bases include, by way ofexample, tertiary amines, such as triethylamine, diisopropylethylamine,N-methylmorpholine and the like. Alternatively, the reaction can beconducted under Schotten-Baumann-type conditions using aqueous alkali,such as sodium hydroxide and the like, as the base. Upon completion ofthe reaction, the resulting sulfonamide 5 is recovered by conventionalmethods including neutralization, extraction, precipitation,chromatography, filtration, and the like.

Other heteroaryl intermediates may also be employed in the abovedescribed reactions including, but not limited to,2-chloro-3-nitropyrazine (J. Med. Chem. 1984, 27, 1634);4-chloro-5-nitroimidazole (J. Chem. Soc. 1930, 268); and the like.

The amino acid derivatives (a) employed in the above reactions areeither known compounds or compounds that can be prepared from knowncompounds by conventional synthetic procedures. For example, amino acidderivatives can be prepared by reacting an organolithium reagent of theformula HetArCH₂Li wherein HetAr group is as defined in the Summary ofthe Invention with commercially available diethyloxalate (Aldrich,Milwaukee, Wis., USA). This reaction is typically conducted by treatingthe diethyloxalate with at least one equivalent of HetArCH₂Li in drypolar organic solvent such as tetrahydrofuran at <−70° C., preferably,−78° C. After addition is completed, the reaction mixture is thenallowed to warm to room temperature and stirred for about 18 to about 20hours. The resulting 3-(HetAr)-oxalate ester is then converted to thecorresponding oxime by following the procedure described in Negi et al,Synthesis, 1996, 8, 991. The oxime is then converted to3-(HetAr)-alanine ethyl ester by the procedure described in Keck et al.Syn. Commun., 1979, 281-286.

Alternatively, a 3-(HetAr)alanine methyl ester can be prepared fromN-benzyloxycarbonyldehydroalanine methyl ester by treating it with HetArhalide wherein HetAr is as defined in the Summary of the Invention, toprovide 3-(HetAr)alaninedehydroalanine methyl ester as described inCarlstrom and Frejd, J. Org. Chem., 1991, 56, 1289-1293. The3-(HetAr)alanine-dehydroalanine methyl ester is then converted to thedesired 3-(HetAr)-alanine methyl ester by the procedure described inKeck et al. Syn. Commun., 1979, 281-286.

Examples of amino acid derivatives suitable for use in the abovereactions include, but are not limited to, L-(N-methyl)histidine methylester, L-(N-benzyl)histidine methyl ester, L-tryptophan methyl ester,β-(2-pyridyl)-L-alanine methyl ester, β-(3-pyridyl)-L-alanine methylester, β-(4-pyridyl)-L-alanine methyl ester, β-(2-thiazolyl)-D,L-alaninemethyl ester, β-(1,2,4-triazol-3-yl)-D,L-alanine methyl ester,β-[2-(2,6-dimethoxyphenyl)pyridyl)-D,L-alanine methyl ester,β-[2-(2,6-dimethoxyphenyl)pyridazinyl)-D,L-alanine methyl ester,β-[2-(2,6-dimethoxyphenyl)pyrimidinyl)-D,L-alanine methyl ester,β-[2-(2,6-dimethoxyphenyl)pyrazinyl)-D,L-alanine methyl ester, and thelike. If desired, of course, other esters or amides of theabove-described compounds may also be employed. Synthesis of compoundssuch as β-[2-(2,6-dimethoxyphenyl)pyridyl)-D,L-alanine methyl ester,β-[2-(2,6-dimethoxyphenyl)pyridazinyl)-D, L-alanine methyl ester,β-[2-(2,6-dimethoxyphenyl)pyrimidinyl)-D,L-alanine methyl ester, andβ-[2-(2,6-dimethoxyphenyl)pyrazinyl)-D,L-alanine methyl ester isdescribed in more detail below.

The sulfonyl chlorides employed in the above reaction are also eitherknown compounds or compounds that can be prepared from known compoundsby conventional synthetic procedures. Such compounds are typicallyprepared from the corresponding sulfonic acid, i.e., from compounds ofthe formula R⁵—SO₃H where R⁵ is as defined above, using phosphoroustrichloride and phosphorous pentachloride. This reaction is generallyconducted by contacting the sulfonic acid with about 2 to 5 molarequivalents of phosphorous trichloride and phosphorous pentachloride,either neat or in an inert solvent, such as dichloromethane, attemperature in the range of about 0° C. to about 80° C. for about 1 toabout 48 hours to afford the sulfonyl chloride. Alternatively, thesulfonyl chloride can be prepared from the corresponding thiol compound,i.e., from compounds of the formula R⁵—SH where R⁵ is as defined herein,by treating the thiol with chlorine (Cl₂) and water under conventionalreaction conditions.

Examples of sulfonyl chlorides suitable for use in this inventioninclude, but are not limited to, methanesulfonyl chloride,2-propanesulfonyl chloride, 1-butanesulfonyl chloride, benzenesulfonylchloride, 1-naphthalene-sulfonyl chloride, 2-naphthalenesulfonylchloride, p-toluenesulfonyl chloride, α-toluenesulfonyl chloride,4-acetamidobenzene-sulfonyl chloride, 4-amidinobenzenesulfonyl chloride,4-tert-butyl-benzenesulfonyl chloride, 4-bromobenzenesulfonyl chloride,2-arboxybenzenesulfonyl chloride, 4-cyanobenzenesulfonyl chloride,3,4-dichlorobenzenesulfonyl chloride, 3,5-dichlorobenzenesulfonylchloride, 3,4-dimethoxybenzenesulfonyl chloride,3,5-ditrifluoromethylbenzenesulfonyl chloride, 4-fluorobenzenesulfonylchloride, 4-methoxybenzenesulfonyl chloride,2-methoxycarbonyl-benzenesulfonyl chloride, 4-methylamidobenzenesulfonylchloride, 4-nitrobenzenesulfonyl chloride, 4-thioamidobenzenesulfonylchloride, 4-trifluoromethylbenzenesulfonyl chloride,4-trifluoromethoxybenzenesulfonyl chloride,2,4,6-trimethylbenzenesulfonyl chloride, 2-phenylethanesulfonylchloride, 2-thiophenesulfonyl chloride, 5-chloro-2-thiophenesulfonylchloride, 2,5-dichloro-4-thiophenesulfonyl chloride, 2-thiazolesulfonylchloride, -methyl-4-thiazolesulfonyl chloride,1-methyl-4-imidazolesulfonyl chloride, 1-methyl-4-pyrazolesulfonylchloride, 5-chloro-1,3dimethyl-4-pyrazole-sulfonyl chloride,3-pyridinesulfonyl chloride, 2-pyrimidinesulfonyl chloride and the like.If desired, a sulfonyl fluoride, sulfonyl bromide or sulfonic acidanhydride may be used in place of the sulfonyl chloride in the abovereaction to form the sulfonamide intermediate 5.

If desired, sulfonamide intermediate 5 can be alkylated at thesulfonamide nitrogen atom to provide compound 6. For example, 5 can becontacted with excess diazomethane (generated, for example, using1-methyl-3-nitro-1-nitrosoguanidine and sodium hydroxide) to afford 6where R⁶ is methyl. Other conventional alkylation procedures andreagents may also be employed to prepare various compounds of thisinvention.

In another preferred embodiment, compounds of this invention may beprepared by displacement of a leaving group as shown in Scheme 2:

where HetAr and X are as defined herein; A′ is heteroaryl, substitutedheteroaryl, heterocyclic or substituted heterocyclic containing twonitrogen atoms in the heteroaryl or heterocyclic ring; and L¹ is aleaving group, such as chloro, bromo, iodo, sulfonate ester and thelike.

Typically, this reaction is conducted by combining approximatelystoichiometric equivalents of 7 and 8 in a suitable inert diluent suchas water, dimethylsulfoxide (DMSO) and the like, with an excess of asuitable base such as sodium bicarbonate, sodium hydroxide, etc. toscavenge the acid generated by the reaction. The reaction is preferablyconducted at from about 25° C. to about 100° C. until reactioncompletion which typically occurs within 1 to about 24 hours. Thisreaction is further described in U.S. Pat. No. 3,598,859, which isincorporated herein by reference in its entirety. Upon reactioncompletion, the product 9 is recovered by conventional methods includingprecipitation, chromatography, filtration and the like.

In still another alternative embodiment, compounds of this invention canbe prepared by reductive amination of a suitable 2-oxocarboxylic acidester, 10, such as a pyruvate ester, as shown in Scheme 3:

where A′, HetAr and X are as defined herein.

Generally, this reaction is conducted by combining equimolar amounts of10 and 11 in an inert diluent such as methanol, ethanol and the likeunder conditions which provide for imine formation (not shown). Theimine formed is then reduced under conventional conditions by a suitablereducing agent such as sodium cyanoborohydride, H₂/palladium on carbonand the like to form the product 12. In a particularly preferredembodiment, the reducing agent is H₂/palladium on carbon which isincorporated into the initial reaction medium thereby permitting iminereduction in situ in a one pot procedure to provide 12. The reaction ispreferably conducted at from about 20° C. to about 80° C. at a pressureof from 1 to 10 atmospheres until reaction completion which typicallyoccurs within 1 to about 24 hours. Upon reaction completion, the product12 is recovered by conventional methods including chromatography,filtration and the like.

Alternatively, certain compounds of this invention can be prepared via arhodium-catalyzed insertion reaction as shown in Scheme 4:

where A″ is heteroaryl or substituted heteroaryl containing two nitrogenatoms in the heteroaryl ring, and HetAr and X (preferably alkoxy) are asdefined herein. Typically, this reaction is conducted using rhodiumacetate dimer, Rh₂(OAc)₄, in an inert diluent such as toluene at atemperature ranging from about 25° C. to about 80° C. for about 1 to 12hours to afford 15. This reaction is described further in B. R. Henkeet. al., J. Med. Chem. 1998, 41, 5020-5036 and references cited therein.

Similarly, certain compounds of this invention can be prepared by thecopper-catalyzed coupling reaction shown in Scheme 5:

where A″ is as defined herein, X³ is halogen, such as chloro, bromo oriodo (preferably iodo), and HetAr and X (preferably alkoxy) are asdefined herein. Typically, this reaction is conducted using copperiodide (CuI) and potassium carbonate in an inert diluent such asN,N-dimethyl acetamide (DMA) at a temperature ranging from about 60° C.to about 120° C. for about 12 to 36 hours to afford 15. This reaction isdescribed further in D. Ma et. al., J. Am. Chem. Soc. 1998, 120,12459-12467 and references cited therein.

For ease of synthesis, the compounds of this invention are typicallyprepared as an ester, i.e., where X is an alkoxy or substituted alkoxygroup and the like. If desired, the ester group can be hydrolysed usingconventional conditions and reagents to provide the correspondingcarboxylic acid. Typically, this reaction is conducted by treating theester with at least one equivalent of an alkali metal hydroxide, such aslithium, sodium or potassium hydroxide, in an inert diluent, such asmethanol or mixtures of methanol and water, at a temperature rangingabout 0° C. to about 24° C. for about 1 to about 12 hours.Alternatively, benzyl esters may be removed by hydrogenolysis using apalladium catalyst, such as palladium on carbon, and tert-butyl esterscan be removed using formic acid to afford the corresponding carboxylicacid.

As will be apparent to those skilled in the art, other functional groupspresent on any of the substituents of the compounds of formulas I-II canbe readily modified or derivatized either before or after theabove-described synthetic reactions using well-known syntheticprocedures. For example, a nitro group present on a substituent of acompound of Formula I-II or an intermediate thereof may be readilyreduced by hydrogenation in the presence of a palladium catalyst, suchas palladium on carbon, to provide the corresponding amino group. Thisreaction is typically conducted at a temperature of from about 20° C. toabout 50° C. for about 6 to about 24 hours in an inert diluent, such asmethanol.

Similarly, a pyridyl group can be hydrogenated in the presence of aplatinum catalyst, such as platinum oxide, in an acidic diluent toprovide the corresponding piperidinyl analogue. Generally, this reactionis conducted by treating the pyridine compound with hydrogen at apressure ranging from about 20 psi to about 60 psi, preferably about 40psi, in the presence of the catalyst at a temperature of about 20° C. toabout 50° C. for about 2 to about 24 hours in an acidic diluent, such asa mixture of methanol and aqueous hydrochloric acid.

Additionally, when the HetAr substituent of a compound of Formula I-IIor an intermediate thereof contains a primary or secondary amino group,such amino groups can be further derivatized either before or after theabove coupling reactions to provide, by way of example, amides,sulfonamides, ureas, thioureas, carbamates, secondary or tertiary aminesand the like. Compounds having a primary amino group on the HetArsubstituent may be prepared, for example, by reduction of thecorresponding nitro compound as described above.

By way of illustration, a compound of Formula I-II or an intermediatethereof having a substituent containing a primary or secondary aminogroup, such as where HetAr is a (4-aminopyridin-2-yl)methyl group, canbe readily N-acylated using conventional acylating reagents andconditions to provide the corresponding amide. This acylation reactionis typically conducted by treating the amino compound with at least oneequivalent, preferably about 1.1 to about 1.2 equivalents, of acarboxylic acid in the presence of a coupling reagent such as acarbodiimide, BOP reagent(benzotriazol-1-yloxy-tris(dimethylamino)-phosphoniumhexafluorophosphonate) and the like, in an inert diluent, such asdichloromethane, chloroform, acetonitrile, tetrahydrofuran,N,N-dimethylformamide and the like, at a temperature ranging from about0° C. to about 37° C. for about 4 to about 24 hours. Preferably, apromoter, such as N-hydroxysuccinimide, 1-hydroxy-benzotriazole and thelike, is used to facilitate the acylation reaction. Examples ofcarboxylic acids suitable for use in this reaction include, but are notlimited to, N-tert-butyloxycarbonylglycine,N-tert-butyloxycarbonyl-L-phenylalanine,N-tert-butyloxycarbonyl-L-aspartic acid benzyl ester, benzoic acid,N-tert-butyloxycarbonylisonipecotic acid, N-methylisonipecotic acid,N-tert-butyloxycarbonylnipecotic acid,N-tert-butyloxycarbonyl-L-tetrahydroisoquinoline-3-carboxylic acid,N-(toluene4-sulfonyl)-L-proline and the like.

Alternatively, a compound of Formula I-II or an intermediate thereofcontaining a primary or secondary amino group can be N-acylated using anacyl halide or a carboxylic acid anhydride to form the correspondingamide. This reaction is typically conducted by contacting the aminocompound with at least one equivalent, preferably about 1.1 to about 1.2equivalents, of the acyl halide or carboxylic acid anhydride in an inertdiluent, such as dichloromethane, at a temperature ranging from about−70° C. to about 40° C. for about 1 to about 24 hours. If desired, anacylation catalyst such as 4-(N,N-dimethylamino)pyridine may be used topromote the acylation reaction. The acylation reaction is preferablyconducted in the presence of a suitable base to scavenge the acidgenerated during the reaction. Suitable bases include, by way ofexample, tertiary amines, such as triethylamine, diisopropylethylamine,N-methylmorpholine and the like. Alternatively, the reaction can beconducted under Schotten-Baumann-type conditions using aqueous alkali,such as sodium hydroxide and the like.

Examples of acyl halides and carboxylic acid anhydrides suitable for usein this reaction include, but are not limited to, 2-methylpropionylchloride, trimethylacetyl chloride, phenylacetyl chloride, benzoylchloride, 2-bromobenzoyl chloride, 2-methylbenzoyl chloride,2-trifluoro-methylbenzoyl chloride, isonicotinoyl chloride, nicotinoylchloride, picolinoyl chloride, acetic anhydride, succinic anhydride, andthe like. Carbamyl chlorides, such as N,N-dimethylcarbamyl chloride,N,N-diethylcarbamyl chloride and the like, can also be used in thisreaction to provide ureas. Similarly, dicarbonates, such asdi-tert-butyl dicarbonate, may be employed to provide carbamates.

In a similar manner, a compound of Formula I-II or an intermediatethereof containing a primary or secondary amino group may beN-sulfonated to form a sulfonamide using a sulfonyl halide or a sulfonicacid anhydride. Sulfonyl halides and sulfonic acid anhydrides suitablefor use in this reaction include, but are not limited to,methanesulfonyl chloride, chloromethane-sulfonyl chloride,p-toluenesulfonyl chloride, trifluoromethanesulfonic anhydride, and thelike. Similarly, sulfamoyl chlorides, such as dimethylsulfamoylchloride, can be used to provide sulfamides (e.g., >N—SO₂—N<).

Additionally, a primary and secondary amino group present on asubstituent of a compound of Formula I-II or an intermediate thereof canbe reacted with an isocyanate or a thioisocyanate to give a urea orthiourea, respectively. This reaction is typically conducted bycontacting the amino compound with at least one equivalent, preferablyabout 1.1 to about 1.2 equivalents, of the isocyanate or thioisocyanatein an inert diluent, such as toluene and the like, at a temperatureranging from about 24° C. to about 37° C. for about 12 to about 24hours. The isocyanates and thioisocyanates used in this reaction arecommercially available or can be prepared from commercially availablecompounds using well-known synthetic procedures. For example,isocyanates and thioisocyanates are readily prepared by reacting theappropriate amine with phosgene or thiophosgene. Examples of isocyanatesand thioisocyanates suitable for use in this reaction include, but arenot limited to, ethyl isocyanate, n-propyl isocyanate, 4-cyanophenylisocyanate, 3-methoxyphenyl isocyanate, 2-phenylethyl isocyanate, methylthioisocyanate, ethyl thioisocyanate, 2-phenylethyl thioisocyanate,3-phenylpropyl thioisocyanate, 3-(N,N-diethylamino)propylthioisocyanate, phenyl thioisocyanate, benzyl thioisocyanate, 3-pyridylthioisocyanate, fluorescein isothiocyanate (isomer I) and the like.

Furthermore, when a compound of Formula I-II or an intermediate thereofcontains a primary or secondary amino group, the amino group can bereductively alkylated using aldehydes or ketones to form a secondary ortertiary amino group. This reaction is typically conducted by contactingthe amino compound with at least one equivalent, preferably about 1.1 toabout 1.5 equivalents, of an aldehyde or ketone and at least oneequivalent based on the amino compound of a metal hydride reducingagent, such as sodium cyanoborohydride, in an inert diluent, such asmethanol, tetrahydrofuran, mixtures thereof and the like, at atemperature ranging from about 0° C. to about 50° C. for about 1 toabout 72 hours. Aldehydes and ketones suitable for use in this reactioninclude, by way of example, benzaldehyde, 4-chlorobenzaldehyde,valeraldehyde and the like.

In a similar manner, when a compound of Formula I-II or an intermediatethereof has a substituent containing a hydroxyl group, the hydroxylgroup can be further modified or derivatized either before or after theabove coupling reactions to provide, by way of example, ethers,carbamates and the like. Compounds having a hydroxyl group on the HetArsubstituent, for example, can be prepared using an amino acid derivativederived from typtophan and the like in the above-described reactions.

By way of example, a compound of Formula I-II or an intermediate thereofhaving a substituent containing a hydroxyl group, such as where HetAr isa (5-hydroxypyridin-2-yl)methyl group, can be readily O-alkylated toform ethers. This O-alkylation reaction is typically conducted bycontacting the hydroxy compound with a suitable alkali or alkaline earthmetal base, such as potassium carbonate, in an inert diluent, such asacetone, 2-butanone and the like, to form the alkali or alkaline earthmetal salt of the hydroxyl group. This salt is generally not isolated,but is reacted in situ with at least one equivalent of an alkyl orsubstituted alkyl halide or sulfonate, such as an alkyl chloride,bromide, iodide, mesylate or tosylate, to afford the ether. Generally,this reaction is conducted at a temperature ranging from about 60° C. toabout 150° C. for about 24 to about 72 hours. Preferably, a catalyticamount of sodium or potassium iodide is added to the reaction mixturewhen an alkyl chloride or bromide is employed in the reaction.

Examples of alkyl or substituted alkyl halides and sulfonates suitablefor use in this reaction include, but are not limited to, tert-butylbromoacetate, N-tert-butyl chloroacetamide, 1-bromoethylbenzene, ethylα-bromophenylacetate, 2-(N-ethyl-N-phenylamino)ethyl chloride,2-(N,N-ethylamino)ethyl chloride, 2-(N,N-diisopropylamino)ethylchloride, 2-(N,N-dibenzylamino)ethyl chloride, 3-(N,N-ethylamino)propylchloride, 3-(N-benzyl-N-methylamino)propyl chloride,N-(2-chloroethyl)morpholine, 2-(hexamethyleneimino)ethyl chloride,3-(N-methylpiperazine)propyl chloride,1-(3-chlorophenyl)-4-(3-chloropropyl)piperazine,2-(4-hydroxy-4-phenylpiperidine)ethyl chloride,N-tert-butyloxycarbonyl-3-piperidinemethyl tosylate, and the like.

Alternatively, a hydroxyl group present on a substituent of a compoundof Formula I-II or an intermediate thereof can be O-alkylating using theMitsunobu reaction. In this reaction, an alcohol, such as3-(N,N-dimethylamino)-1-propanol and the like, is reacted with about 1.0to about 1.3 equivalents of triphenylphosphine and about 1.0 to about1.3 equivalents of diethyl azodicarboxylate in an inert diluent, such astetrahydrofuran, at a temperature ranging from about −10° C. to about 5°C. for about 0.25 to about 1 hour. About 1.0 to about 1.3 equivalents ofa hydroxy compound, such as N-tert-butyltyrosine methyl ester, is thenadded and the reaction mixture is stirred at a temperature of about 0°C. to about 30° C. for about 2 to about 48 hours to provide theO-alkylated product.

In a similar manner, a compound of Formula I-II or an intermediatethereof containing an aryl or heteroaryl hydroxy group can be reactedwith an aryl iodide to provide a diaryl ether or a -heteroaryl-O-arylether. Generally, this reaction is conducted by forming the alkali metalsalt of the hydroxyl group using a suitable base, such as sodiumhydride, in an inert diluent such as xylenes at a temperature of about−25° C. to about 10° C. The salt is then treated with about 1.1 to about1.5 equivalents of cuprous bromide dimethyl sulfide complex at atemperature ranging from about 10° C. to about 30° C. for about 0.5 toabout 2.0 hours, followed by about 1.1 to about 1.5 equivalents of anaryl iodide, such as sodium 2-iodobenzoate and the like. The reaction isthen heated to about 70° C. to about 150° C. for about 2 to about 24hours to provide the diaryl ether or the -heteroaryl-O-aryl ether.

Additionally, a hydroxy-containing compound can also be readilyderivatized to form a carbamate. In one method for preparing suchcarbamates, a hydroxy compound of Formula I-II or an intermediatethereof is contacted with about 1.0 to about 1.2 equivalents of4-nitrophenyl chloroformate in an inert diluent, such asdichloromethane, at a temperature ranging from about −25° C. to about 0°C. for about 0.5 to about 2.0 hours. Treatment of the resultingcarbonate with an excess, preferably about 2 to about 5 equivalents, ofa trialkylamine, such as triethylamine, for about 0.5 to 2 hours,followed by about 1.0 to about 1.5 equivalents of a primary or secondaryamine provides the carbamate. Examples of amines suitable for use inthis reaction include, but are not limited to, piperazine,1-methylpiperazine, 1-acetylpiperazine, morpholine, thiomorpholine,pyrrolidine, piperidine, N,N-dimethylamine and the like.

Alternatively, in another method for preparing carbamates, ahydroxy-containing compound is contacted with about 1.0 to about 1.5equivalents of a carbamyl chloride in an inert diluent, such asdichloromethane, at a temperature ranging from about 25° C. to about 70°C. for about 2 to about 72 hours. Typically, this reaction is conductedin the presence of a suitable base to scavenge the acid generated duringthe reaction. Suitable bases include, by way of example, tertiaryamines, such as triethylamine, diisopropylethylamine, N-methylmorpholineand the like. Additionally, at least one equivalent (based on thehydroxy compound) of 4-(N,N-dimethylamino)pyridine is preferably addedto the reaction mixture to facilitate the reaction. Examples of carbamylchlorides suitable for use in this reaction include, by way of example,dimethylcarbamyl chloride, diethylcarbamyl chloride and the like.

Likewise, when a compound of Formula I-II or an intermediate thereofcontains a primary or secondary hydroxyl group, such hydroxyl groups canbe readily converted into a leaving group and displaced to form, forexample, amines, sulfides and fluorides. Generally, when a chiralcompound is employed in these reactions, the stereochemistry at thecarbon atom attached to the derivatized hydroxyl group is typicallyinverted.

These reactions are typically conducted by first converting the hydroxylgroup into a leaving group, such as a tosylate, by treatment of thehydroxy compound with at least one equivalent of a sulfonyl halide, suchas p-toluenesulfonyl chloride and the like, in pyridine. This reactionis generally conducted at a temperature of from about 0° C. to about 70°C. for about 1 to about 48 hours. The resulting tosylate can then bereadily displaced with sodium azide, for example, by contacting thetosylate with at least one equivalent of sodium azide in an inertdiluent, such as a mixture of N,N-dimethylformamide and water, at atemperature ranging from about 0° C. to about 37° C. for about 1 toabout 12 hours to provide the corresponding azido compound. The azidogroup can then be reduced by, for example, hydrogenation using apalladium on carbon catalyst to provide the amino (—NH₂) compound.

Similarly, a tosylate group can be readily displaced by a thiol to forma sulfide. This reaction is typically conducted by contacting thetosylate with at least one equivalent of a thiol, such as thiophenol, inthe presence of a suitable base, such as1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), in an inert diluent, such asN,N-dimethylformamide, at a temperature of from about 0° C. to about 37°C. for about 1 to about 12 hours to provide the sulfide. Additionally,treatment of a tosylate with morpholinosulfur trifluoride in an inertdiluent, such as dichloromethane, at a temperature ranging from about 0°C. to about 37° C. for about 12 to about 24 hours affords thecorresponding fluoro compound.

Furthermore, a compound of Formula I-II or an intermediate thereofhaving a substituent containing an iodoheteroaryl group, for example,when HetAr is a (4-iodopyridyl)methyl group, can be readily convertedeither before or after the above coupling reactions into a-heteroaryl-aryl compound. Typically, this reaction is conducted bytreating the iodoheteroaryl compound with about 1.1 to about 2equivalents of an arylzinc iodide, such as 2-(methoxycarbonyl)phenylzinciodide, in the presence of a palladium catalyst, such as palladiumtetra(triphenylphosphine), in an inert diluent, such as tetrahydrofuran,at a temperature ranging from about 24° C. to about 30° C. untilreaction completion. This reaction is further described, for example, inRieke, J. Org. Chem. 1991, 56, 1445. Additional methods for preparing-heteroaryl-aryl derivatives are disclosed in International PublicationNumber WO 98/53817, published Dec. 3, 1998, the disclosure of which isincorporated herein by reference in its entirety. The -heteroaryl-arylcompound can also be prepared by coupling (4-iodopyridyl)methyl groupwith arylboronic acid under Suzuki coupling reaction conditions wellknown in the art.

In some cases, the compounds of Formula I-II or intermediates thereofmay contain substituents having one or more sulfur atoms. When present,such sulfur atoms can be oxidized either before or after the abovecoupling reactions to provide a sulfoxide or sulfone compound usingconventional reagents and reaction conditions. Suitable reagents foroxidizing a sulfide compound to a sulfoxide include, by way of example,hydrogen peroxide, 3-chloroperoxy-benzoic acid (MCPBA), sodium periodateand the like. The oxidation reaction is typically conducted bycontacting the sulfide compound with about 0.95 to about 1.1 equivalentsof the oxidizing reagent in an inert diluent, such as dichloromethane,at a temperature ranging from about −50° C. to about 75° C. for about 1to about 24 hours. The resulting sulfoxide can then be further oxidizedto the corresponding sulfone by contacting the sulfoxide with at leastone additional equivalent of an oxidizing reagent, such as hydrogenperoxide, MCPBA, potassium permanganate and the like. Alternatively, thesulfone can be prepared directly by contacting the sulfide with at leasttwo equivalents, and preferably an excess, of the oxidizing reagent.Such reactions are described further in March, “Advanced OrganicChemistry”, 4th Ed., pp. 1201-1202, Wiley Publisher, 1992.

Other procedures and reaction conditions for preparing the compounds ofthis invention are described in the examples set forth below.Additionally, other procedures for preparing compounds useful in certainaspects of this invention are disclosed in U.S. applications Ser. Nos.09/489,377 and 09/489,378, filed on Jan. 21, 2000, entitled “CompoundsWhich Inhibit Leucocyte Adhesion Mediated by VLA-4” (Attorney DocketNos. 002010-517 and 002010-525); the disclosures of which areincorporated herein by reference in their entirety.

Pharmaceutical Formulations

When employed as pharmaceuticals, the compounds of this invention areusually administered in the form of pharmaceutical compositions. Thesecompounds can be administered by a variety of routes including oral,rectal, transdermal, subcutaneous, intravenous, intramuscular, andintranasal. These compounds are effective as both injectable and oralcompositions. Such compositions are prepared in a manner well known inthe pharmaceutical art and comprise at least one active compound.

This invention also includes pharmaceutical compositions which contain,as the active ingredient, one or more of the compounds of Formula I-IIabove associated with pharmaceutically acceptable carriers. In makingthe compositions of this invention, the active ingredient is usuallymixed with an excipient, diluted by an excipient or enclosed within sucha carrier which can be in the form of a capsule, sachet, paper or othercontainer. The excipient employed is typically an excipient suitable foradministration to human subjects or other mammals. When the excipientserves as a diluent, it can be a solid, semi-solid, or liquid material,which acts as a vehicle, carrier or medium for the active ingredient.Thus, the compositions can be in the form of tablets, pills, powders,lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions,syrups, aerosols (as a solid or in a liquid medium), ointmentscontaining, for example, up to 10% by weight of the active compound,soft and hard gelatin capsules, suppositories, sterile injectablesolutions, and sterile packaged powders.

In preparing a formulation, it may be necessary to mill the activecompound to provide the appropriate particle size prior to combiningwith the other ingredients. If the active compound is substantiallyinsoluble, it ordinarily is milled to a particle size of less than 200mesh. If the active compound is substantially water soluble, theparticle size is normally adjusted by milling to provide a substantiallyuniform distribution in the formulation, e.g. about 40 mesh.

Some examples of suitable excipients include lactose, dextrose, sucrose,sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates,tragacanth, gelatin, calcium silicate, microcrystalline cellulose,polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose. Theformulations can additionally include: lubricating agents such as talc,magnesium stearate, and mineral oil; wetting agents; emulsifying andsuspending agents; preserving agents such as methyl- andpropylhydroxy-benzoates; sweetening agents; and flavoring agents. Thecompositions of the invention can be formulated so as to provide quick,sustained or delayed release of the active ingredient afteradministration to the patient by employing procedures known in the art.

The compositions are preferably formulated in a unit dosage form, eachdosage containing from about 5 to about 100 mg, more usually about 10 toabout 30 mg, of the active ingredient. The term “unit dosage forms”refers to physically discrete units suitable as unitary dosages forhuman subjects and other mammals, each unit containing a predeterminedquantity of active material calculated to produce the desiredtherapeutic effect, in association with a suitable pharmaceuticalexcipient.

The active compound is effective over a wide dosage range and isgenerally administered in a pharmaceutically effective amount. It, willbe understood, however, that the amount of the compound actuallyadministered will be determined by a physician, in the light of therelevant circumstances, including the condition to be treated, thechosen route of administration, the actual compound administered, theage, weight, and response of the individual patient, the severity of thepatient's symptoms, and the like.

For preparing solid compositions such as tablets, the principal activeingredient is mixed with a pharmaceutical excipient to form a solidpreformulation composition containing a homogeneous mixture of acompound of the present invention. When referring to thesepreformulation compositions as homogeneous, it is meant that the activeingredient is dispersed evenly throughout the composition so that thecomposition may be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules. This solid preformulation isthen subdivided into unit dosage forms of the type described abovecontaining from, for example, 0.1 to about 500 mg of the activeingredient of the present invention.

The tablets or pills of the present invention may be coated or otherwisecompounded to provide a dosage form affording the advantage of prolongedaction. For example, the tablet or pill can comprise an inner dosage andan outer dosage component, the latter being in the form of an envelopeover the former. The two components can be separated by an enteric layerwhich serves to resist disintegration in the stomach and permit theinner component to pass intact into the duodenum or to be delayed inrelease. A variety of materials can be used for such enteric layers orcoatings, such materials including a number of polymeric acids andmixtures of polymeric acids with such materials as shellac, cetylalcohol, and cellulose acetate.

The liquid forms in which the novel compositions of the presentinvention may be incorporated for administration orally or by injectioninclude aqueous solutions suitably flavored syrups, aqueous or oilsuspensions, and flavored emulsions with edible oils such as cottonseedoil, sesame oil, coconut oil, or peanut oil, as well as elixirs andsimilar pharmaceutical vehicles.

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically acceptable excipients as describedsupra. Preferably the compositions are administered by the oral or nasalrespiratory route for local or systemic effect. Compositions inpreferably pharmaceutically acceptable solvents may be nebulized by useof inert gases. Nebulized solutions may be breathed directly from thenebulizing device or the nebulizing device may be attached to a facemasks tent, or intermittent positive pressure breathing machine.Solution, suspension, or powder compositions may be administered,preferably orally or nasally, from devices which deliver the formulationin an appropriate manner.

The following formulation examples illustrate the pharmaceuticalcompositions of the present invention.

FORMULATION EXAMPLE 1

Hard gelatin capsules containing the following ingredients are prepared:

Quantity Ingredient (mg/capsule) Active Ingredient 30.0 Starch 305.0Magnesium stearate 5.0

The above ingredients are mixed and filled into hard gelatin capsules in340 mg quantities.

FORMULATION EXAMPLE 2

A tablet formula is prepared using the ingredients below:

Quantity Ingredient (mg/tablet) Active Ingredient 25.0 Cellulose,microcrystalline 200.0 Colloidal silicon dioxide 10.0 Stearic acid 5.0

The components are blended and compressed to form tablets, each weighing240 mg.

FORMULATION EXAMPLE 3

A dry powder inhaler formulation is prepared containing the followingcomponents:

Ingredient Weight % Active Ingredient 5 Lactose 95

The active mixture is mixed with the lactose and the mixture is added toa dry powder inhaling appliance.

FORMULATION EXAMPLE 4

Tablets, each containing 30 mg of active ingredient, are prepared asfollows:

Quantity Ingredient (mg/tablet) Active Ingredient 30.0 mg Starch 45.0 mgMicrocrystalline cellulose 35.0 mg Polyvinylpyrrolidone  4.0 mg (as 10%solution in water) Sodium carboxymethyl starch  4.5 mg Magnesiumstearate  0.5 mg Talc  1.0 mg Total  120 mg

The active ingredient, starch and cellulose are passed through a No. 20mesh U.S. sieve and mixed thoroughly. The solution ofpolyvinyl-pyrrolidone is mixed with the resultant powders, which arethen passed through a 16 mesh U.S. sieve. The granules so produced aredried at 50° to 60° C. and passed through a 16 mesh U.S. sieve. Thesodium carboxymethyl starch, magnesium stearate, and talc, previouslypassed through a No. 30 mesh U.S. sieve, are then added to the granuleswhich, after mixing, are compressed on a tablet machine to yield tabletseach weighing 150 mg.

FORMULATION EXAMPLE 5

Capsules, each containing 40 mg of medicament are made as follows:

Quantity Ingredient (mg/capsule) Active Ingredient  40.0 mg Starch 109.0mg Magnesium stearate  1.0 mg Total 150.0 mg

The active ingredient, cellulose, starch, an magnesium stearate areblended, passed through a No. 20 mesh U.S. sieve, and filled into hardgelatin capsules in 150 mg quantities.

FORMULATION EXAMPLE 6

Suppositories, each containing 25 mg of active ingredient are made asfollows:

Ingredient Amount Active Ingredient   25 mg Saturated fatty acidglycerides to 2,000 mg

The active ingredient is passed through a No. 60 mesh U.S. sieve andsuspended in the saturated fatty acid glycerides previously melted usingthe minimum heat necessary. The mixture is then poured into asuppository mold of nominal 2.0 g capacity and allowed to cool.

FORMULATION EXAMPLE 7

Suspensions, each containing 50 mg of medicament per 5.0 ml dose aremade as follows:

Ingredient Amount Active Ingredient 50.0 mg Xanthan gum  4.0 mg Sodiumcarboxymethyl cellulose (11%) 50.0 mg Microcrystalline cellulose (89%)Sucrose 1.75 g  Sodium benzoate 10.0 mg Flavor and Color q.v. Purifiedwater to  5.0 ml

The medicament, sucrose and xanthan gum are blended, passed through aNo. 10 mesh U.S. sieve, and then mixed with a previously made solutionof the microcrystalline cellulose and sodium carboxymethyl cellulose inwater. The sodium benzoate, flavor, and color are diluted with some ofthe water and added with stirring. Sufficient water is then added toproduce the required volume.

FORMULATION EXAMPLE 8

Quantity Ingredient (mg/capsule) Active Ingredient  15.0 mg Starch 407.0mg Magnesium stearate  3.0 mg Total 425.0 mg

The active ingredient, cellulose, starch, and magnesium stearate areblended, passed through a No. 20 mesh U.S. sieve, and filled into hardgelatin capsules in 560 mg quantities.

FORMULATION EXAMPLE 9

An intravenous formulation may be prepared as follows:

Ingredient Quantity Active Ingredient 250.0 mg Isotonic saline  1000 mL

FORMULATION EXAMPLE 10

A topical formulation may be prepared as follows:

Ingredient Quantity Active Ingredient 1-10 g Emulsifying Wax 30 g LiquidParaffin 20 g White Soft Paraffin to 100 g

The white soft paraffin is heated until molten. The liquid paraffin andemulsifing wax are incorporated an stirred until dissolved. The activeingredient is added and stirring is continued until dispersed. Themixture is then cooled until solid.

Another preferred formulation employed in the methods of the presentinvention employs transdermal delivery devices (“patches”). Suchtransdermal patches may be used to provide continuous or discontinuousinfusion of the compounds of the present invention in controlledamounts. The construction and use of transdermal patches for thedelivery of pharmaceutical agents is well known in the art. See, e.g.,U.S. Pat. No. 5,023,252, issued Jun. 11, 1991, herein incorporated byreference. Such patches may be constructed for continuous, pulsatile, oron demand delivery of pharmaceutical agents.

Direct or indirect placement techniques may be used when it is desirableor necessary to introduce the pharmaceutical composition to the brain.Direct techniques usually involve placement of a drug delivery catheterinto the host's ventricular system to bypass the blood-brain barrier.One such implantable delivery system used for the transport ofbiological factors to specific anatomical regions of the body isdescribed in U.S. Pat. No. 5,011,472 which is herein incorporated byreference.

Indirect techniques, which are generally preferred, usually involveformulating the compositions to provide for drug latentiation by theconversion of hydrophilic drugs into lipid-soluble drugs. Latentiationis generally achieved through blocking of the hydroxy, carbonyl,sulfate, and primary amine groups present on the drug to render the drugmore lipid soluble and amenable to transportation across the blood-brainbarrier. Alternatively, the delivery of hydrophilic drugs may beenhanced by intra-arterial infusion of hypertonic solutions which cantransiently open the blood-brain barrier.

Utility

The compounds of this invention can be employed to bind VLA-4 (α₄β₁integrin) in biological samples, i.e., the compounds bind VLA-4 with anIC₅₀ of 15 μM or less in a competitive binding assay as describedherein. Accordingly, these compounds have utility in, for example,assaying such samples for VLA-4. In such assays, the compounds can bebound to a solid support and the VLA-4 sample added thereto. The amountof VLA-4 in the sample can be determined by conventional methods such asuse of a sandwich ELISA assay. Alternatively, labeled VLA-4 can be usedin a competitive assay to measure for the presence of VLA-4 in thesample. Other suitable assays are well known in the art.

In addition, certain of the compounds of this invention inhibit, invivo, adhesion of leukocytes to endothelial cells mediated by VLA-4 bycompetitive binding to VLA-4. Accordingly, the compounds of thisinvention can be used in the treatment of diseases mediated by VLA-4 orleucocyte adhesion. Such diseases include inflammatory diseases inmammalian patients such as asthma, Alzheimer's disease, atherosclerosis,AIDS dementia, diabetes (including acute juvenile onset diabetes),inflammatory bowel disease (including ulcerative colitis and Crohn'sdisease), multiple sclerosis, rheumatoid arthritis, tissuetransplantation, tumor metastasis, meningitis, encephalitis, stroke, andother cerebral traumas, nephritis, retinitis, atopic dermatitis,psoriasis, myocardial ischemia and acute leukocyte-mediated lung injurysuch as that which occurs in adult respiratory distress syndrome.

The biological activity of the compounds identified above may be assayedin a variety of systems. For example, a compound can be immobilized on asolid surface and adhesion of cells expressing VLA-4 can be measured.Using such formats, large numbers of compounds can be screened. Cellssuitable for this assay include any leukocytes known to express VLA-4such as T cells, B cells, monocytes, eosinophils, and basophils. Anumber of leukocyte cell lines can also be used, examples include Jurkatand U937.

The test compounds can also be tested for the ability to competitivelyinhibit binding between VLA-4 and VCAM-1, or between VLA-4 and a labeledcompound known to bind VLA-4 such as a compound of this invention orantibodies to VLA-4. In these assays, the VCAM-1 can be immobilized on asolid surface. VCAM-1 may also be expressed as a recombinant fusionprotein having an Ig tail (e.g., IgG) so that binding to VLA-4 may bedetected in an immunoassay. Alternatively, VCAM-1 expressing cells, suchas activated endothelial cells or VCAM-1 transfected fibroblasts can beused. For assays to measure the ability to block adhesion to brainendothelial cells, the assays described in International PatentApplication Publication No. WO 91/05038 are particularly preferred. Thisapplication is incorporated herein by reference in its entirety.

Many assay formats employ labeled assay components. The labeling systemscan be in a variety of forms. The label may be coupled directly orindirectly to the desired component of the assay according to methodswell known in the art. A wide variety of labels may be used. Thecomponent may be labelled by any one of several methods. The most commonmethod of detection is the use of autoradiography with ³H, ¹²⁵I, ³⁵S,¹⁴C, or ³²P labeled compounds or the like. Non-radioactive labelsinclude ligands which bind to labeled antibodies, fluorophores,chemiluminescent agents, enzymes and antibodies which can serve asspecific binding pair members for a labeled ligand. The choice of labeldepends on sensitivity required, ease of conjugation with the compound,stability requirements, and available instrumentation.

Appropriate in vivo models for demonstrating efficacy in treatinginflammatory responses include EAE (experimental autoimmuneencephalomyelitis) in mice, rats, guinea pigs or primates, as well asother inflammatory models dependent upon α4 integrins.

Compounds having the desired biological activity may be modified asnecessary to provide desired properties such as improved pharmacologicalproperties (e.g., in vivo stability, bio-availability), or the abilityto be detected in diagnostic applications. Stability can be assayed in avariety of ways such as by measuring the half-life of the proteinsduring incubation with peptidases or human plasma or serum. A number ofsuch protein stability assays have been described (see, e.g., Verhoef etal., Eur. J. Drug Metab. Pharmacokinet., 1990, 15(2):83-93).

For diagnostic purposes, a wide variety of labels may be linked to thecompounds, which may provide, directly or indirectly, a detectablesignal. Thus, the compounds of the subject invention may be modified ina variety of ways for a variety of end purposes while still retainingbiological activity. In addition, various reactive sites may beintroduced at the terminus for linking to particles, solid substrates,macromolecules, or the like.

Labeled compounds can be used in a variety of in vivo or in vitroapplications. A wide variety of labels may be employed, such asradionuclides (e.g., gamma-emitting radioisotopes such as technetium-99or indium-111), fluorescers (e.g., fluorescein), enzymes, enzymesubstrates, enzyme cofactors, enzyme inhibitors, chemiluminescentcompounds, bioluminescent compounds, and the like. Those of ordinaryskill in the art will know of other suitable labels for binding to thecomplexes, or will be able to ascertain such using routineexperimentation. The binding of these labels is achieved using standardtechniques common to those of ordinary skill in the art.

In vitro uses include diagnostic applications such as monitoringinflammatory responses by detecting the presence of leukocytesexpressing VLA-4. The compounds of this invention can also be used forisolating or labeling such cells. In addition, as mentioned above, thecompounds of the invention can be used to assay for potential inhibitorsof VLA-4/VCAM-1 interactions.

For in vivo diagnostic imaging to identify, e.g., sites of inflammation,radioisotopes are typically used in accordance with well knowntechniques. The radioisotopes may be bound to the peptide eitherdirectly or indirectly using intermediate functional groups. Forinstance, chelating agents such as diethylenetriaminepentacetic acid(DTPA) and ethylenediaminetetraacetic acid (EDTA) and similar moleculeshave been used to bind proteins to metallic ion radioisotopes.

The complexes can also be labeled with a paramagnetic isotope forpurposes of in vivo diagnosis, as in magnetic resonance imaging (MRI) orelectron spin resonance (ESR), both of which are well known. In general,any conventional method for visualizing diagnostic imaging can be used.Usually gamma- and positron-emitting radioisotopes are used for cameraimaging and paramagnetic isotopes are used for MRI. Thus, the compoundscan be used to monitor the course of amelioration of an inflammatoryresponse in an individual. By measuring the increase or decrease inlymphocytes expressing VLA-4 it is possible to determine whether aparticular therapeutic regimen aimed at ameliorating the disease iseffective.

The pharmaceutical compositions of the present invention can be used toblock or inhibit cellular adhesion associated with a number of diseasesand disorders. For instance, a number of inflammatory disorders areassociated with integrins or leukocytes. Treatable disorders include,e.g., transplantation rejection (e.g., allograft rejection), Alzheimer'sdisease, atherosclerosis, AIDS dementia, diabetes (including acutejuvenile onset diabetes), retinitis, cancer metastases, rheumatoidarthritis, acute leukocyte-mediated lung injury (e.g., adult respiratorydistress syndrome), asthma, nephritis, and acute and chronicinflammation, including atopic dermatitis, psoriasis, myocardialischemia, and inflammatory bowel disease (including Crohn's disease andulcerative colitis). In preferred embodiments the pharmaceuticalcompositions are used to treat inflammatory brain disorders, such asmultiple sclerosis (MS), viral meningitis and encephalitis.

Inflammatory bowel disease is a collective term for two similar diseasesreferred to as Crohn's disease and ulcerative colitis. Crohn's diseaseis an idiopathic, chronic ulceroconstrictive inflammatory diseasecharacterized by sharply delimited and typically transmural involvementof all layers of the bowel wall by a granulomatous inflammatoryreaction. Any segment of the gastrointestinal tract, from the mouth tothe anus, may be involved, although the disease most commonly affectsthe terminal ileum and/or colon. Ulcerative colitis is an inflammatoryresponse limited largely to the colonic mucosa and submucosa.Lymphocytes and macrophages are numerous in lesions of inflammatorybowel disease and may contribute to inflammatory injury.

Asthma is a disease characterized by increased responsiveness of thetracheobronchial tree to various stimuli potentiating paroxysmalconstriction of the bronchial airways. The stimuli cause release ofvarious mediators of inflammation from IgE-coated mast cells includinghistamine, eosinophilic and neutrophilic chemotactic factors,leukotrines, prostaglandin and platelet activating factor. Release ofthese factors recruits basophils, eosinophils and neutrophils, whichcause inflammatory injury.

Atherosclerosis is a disease of arteries (e.g., coronary, carotid, aortaand iliac). The basic lesion, the atheroma, consists of a raised focalplaque within the intima, having a core of lipid and a covering fibrouscap. Atheromas compromise arterial blood flow and weaken affectedarteries. Myocardial and cerebral infarcts are a major consequence ofthis disease. Macrophages and leukocytes are recruited to atheromas andcontribute to inflammatory injury.

Rheumatoid arthritis is a chronic, relapsing inflammatory disease thatprimarily causes impairment and destruction of joints. Rheumatoidarthritis usually first affects the small joints of the hands and feetbut then may involve the wrists, elbows, ankles and knees. The arthritisresults from interaction of synovial cells with leukocytes thatinfiltrate from the circulation into the synovial lining of the joints.See e.g., Paul, Immunology (3d ed., Raven Press, 1993).

Another indication for the compounds of this invention is in treatmentof organ or graft rejection mediated by VLA-4. Over recent years therehas been a considerable improvement in the efficiency of surgicaltechniques for transplanting tissues and organs such as skin, kidney,liver, heart, lung, pancreas and bone marrow. Perhaps the principaloutstanding problem is the lack of satisfactory agents for inducingimmunotolerance in the recipient to the transplanted allograft or organ.When allogeneic cells or organs are transplanted into a host (i.e., thedonor and donee are different individuals from the same species), thehost immune system is likely to mount an immune response to foreignantigens in the transplant (host-versus-graft disease) leading todestruction of the transplanted tissue. CD8⁺ cells, CD4 cells andmonocytes are all involved in the rejection of transplant tissues.Compounds of this invention which bind to alpha-4 integrin are useful,inter alia, to block alloantigen-induced immune responses in the doneethereby preventing such cells from participating in the destruction ofthe transplanted tissue or organ. See, e.g., Paul et al., TransplantInternational 9, 420-425 (1996); Georczynski et al., Immunology 87,573-580 (1996); Georcyznski et al., Transplant. Immunol. 3, 55-61(1995); Yang et al., Transplantation 60, 71-76 (1995); Anderson et al.,APMIS 102, 23-27 (1994).

A related use for compounds of this invention which bind to VLA-4 is inmodulating the immune response involved in “graft versus host” disease(GVHD). See e.g., Schlegel et al., J. Immunol. 155, 3856-3865 (1995).GVHD is a potentially fatal disease that occurs when immunologicallycompetent cells are transferred to an allogeneic recipient. In thissituation, the donor's immunocompetent cells may attack tissues in therecipient. Tissues of the skin, gut epithelia and liver are frequenttargets and may be destroyed during the course of GVHD. The diseasepresents an especially severe problem when immune tissue is beingtransplanted, such as in bone marrow transplantation; but less severeGVHD has also been reported in other cases as well, including heart andliver transplants. The therapeutic agents of the present invention areused, inter alia, to block activation of the donor T-cells therebyinterfering with their ability to lyse target cells in the host.

A further use of the compounds of this invention is inhibiting tumormetastasis. Several tumor cells have been reported to express VLA-4 andcompounds which bind VLA-4 block adhesion of such cells to endothelialcells. Steinback et al., Urol. Res. 23, 175-83 (1995); Orosz et al.,Int. J. Cancer 60, 867-71 (1995); Freedman et al., Leuk. Lymphoma 13,47-52 (1994); Okahara et al., Cancer Res. 54, 3233-6 (1994).

A further use of the compounds of this invention is in treating multiplesclerosis. Multiple sclerosis is a progressive neurological autoimmunedisease that affects an estimated 250,000 to 350,000 people in theUnited States. Multiple sclerosis is thought to be the result of aspecific autoimmune reaction in which certain leukocytes attack andinitiate the destruction of myelin, the insulating sheath covering nervefibers. In an animal model for multiple sclerosis, murine monoclonalantibodies directed against VLA-4 have been shown to block the adhesionof leukocytes to the endothelium, and thus prevent inflammation of thecentral nervous system and subsequent paralysis in the animals¹⁶.

Pharmaceutical compositions of the invention are suitable for use in avariety of drug delivery systems. Suitable formulations for use in thepresent invention are found in Remington's Pharmaceutical Sciences, MacePublishing Company, Philadelphia, Pa., 17th ed. (1985).

In order to enhance serum half-life, the compounds may be encapsulated,introduced into the lumen of liposomes, prepared as a colloid, or otherconventional techniques may be employed which provide an extended serumhalf-life of the compounds. A variety of methods are available forpreparing liposomes, as described in, e.g., Szoka, et al., U.S. Pat.Nos. 4,235,871, 4,501,728 and 4,837,028 each of which is incorporatedherein by reference.

The amount administered to the patient will vary depending upon what isbeing administered, the purpose of the administration, such asprophylaxis or therapy, the state of the patient, the manner ofadministration, and the like. In therapeutic applications, compositionsare administered to a patient already suffering from a disease in anamount sufficient to cure or at least partially arrest the symptoms ofthe disease and its complications. An amount adequate to accomplish thisis defined as “therapeutically effective dose.” Amounts effective forthis use will depend on the disease condition being treated as well asby the judgment of the attending clinician depending upon factors suchas the severity of the inflammation, the age, weight and generalcondition of the patient, and the like.

The compositions administered to a patient are in the form ofpharmaceutical compositions described above. These compositions may besterilized by conventional sterilization techniques, or may be sterilefiltered. The resulting aqueous solutions may be packaged for use as is,or lyophilized, the lyophilized preparation being combined with asterile aqueous carrier prior to administration. The pH of the compoundpreparations typically will be between 3 and 11, more preferably from 5to 9 and most preferably from 7 to 8. It will be understood that use ofcertain of the foregoing excipients, carriers, or stabilizers willresult in the formation of pharmaceutical salts.

The therapeutic dosage of the compounds of the present invention willvary according to, for example, the particular use for which thetreatment is made, the manner of administration of the compound, thehealth and condition of the patient, and the judgment of the prescribingphysician. For example, for intravenous administration, the dose willtypically be in the range of about 20 μg to about 500 μg per kilogrambody weight, preferably about 100 μg to about 300 μg per kilogram bodyweight. Suitable dosage ranges for intranasal administration aregenerally about 0.1 pg to 1 mg per kilogram body weight. Effective dosescan be extrapolated from dose-response curves derived from in vitro oranimal model test systems.

Compounds of this invention are also capable of binding or antagonizingthe actions of α₆β₁, α₉β₁, α₄β₇, α_(d)β₂, α_(e)β₇ integrins (althoughα₄β₁ and α₉β₁ are preferred in this invention). Accordingly, compoundsof this invention are also useful for preventing or reversing thesymptoms, disorders or diseases induced by the binding of theseintegrins to their respective ligands.

For example, International Publication Number WO 98/53817, publishedDec. 3, 1998 (the disclosure of which is incorporated herein byreference in its entirety) and references cited therein describedisorders mediated by α₄β₇. This reference also describes an assay fordetermining antagonism of α₄β₇ dependent binding to VCAM-Ig fusionprotein.

Additionally, compounds that bind α_(d)β₂ and α_(e)β₇ integrins areparticularly useful for the treatment of asthma and related lungdiseases. See, for example, M. H. Grayson et al., J. Exp. Med. 1998,188(11) 2187-2191. Compounds that bind α_(e)β₇ integrin are also usefulfor the treatment of systemic lupus erythematosus (see, for example, M.Pang et al., Arthritis Rheum. 1998, 41(8), 1456-1463); Crohn's disease,ulcerative colitis and infammatory bowel disease (IBD) (see, forexample, D. Elewaut et al., Scand J. Gastroenterol 1998, 33(7) 743-748);Sjogren's syndrome (see, for example, U. Kroneld et al., Scand J.Gastroenterol 1998, 27(3), 215-218); and rheumatoid arthritis (see, forexample, Scand J. Gastroenterol 1996, 44(3), 293-298). And compoundsthat bind α₆β₁ may be useful in preventing fertilization (see, forexample, H. Chen et al., Chem. Biol. 1999, 6, 1-10).

Certain of the compounds within the generic formulas described hereinare also useful as synthetic intermediates for other compounds of thisinvention as illustrated in the examples herein.

The following synthetic and biological examples are offered toillustrate this invention and are not to be construed in any way aslimiting the scope of this invention. Unless otherwise stated, alltemperatures are in degrees Celsius.

EXAMPLES

In the examples below, the following abbreviations have the followingmeanings. If an abbreviation is not defined, it has its generallyaccepted meaning.

aq or aq. = aqueous AcOH = acetic acid bd = broad doublet bm = broadmultiplet bs = broad singlet Bn = benzyl Boc = N-tert-butoxylcarbonylBoc₂O = di-tert-butyl dicarbonate BOP = benzotriazol-1-yloxy-tris(dimethylamino)phosphonium hexafluorophosphate Cbz = carbobenzyloxyCHCl₃ = chloroform CH₂Cl₂ = dichloromethane (COCl)₂ = oxalyl chloride d= doublet dd = doublet of doublets dt = doublet of triplets DBU =1,8-diazabicyclo[5.4.0]undec-7-ene DCC = 1,3-dicyclohexylcarbodiimideDMAP = 4-N,N-dimethylaminopyridine DME = ethylene glycol dimethyl etherDMF = N,N-dimethylformamide DMSO = dimethylsulfoxide EDC =1-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride Et₃N =triethylamine Et₂O = diethyl ether EtOAc = ethyl acetate EtOH = ethanoleq or eq. = equivalent Fmoc = N-(9-fluorenylmethoxycarbonyl) FmocONSu =N-(9-fluorenylmethoxycarbonyl)- succinimide g = grams h = hour H₂O =water HBr = hydrobromic acid HCl = hydrochloric acid HOBT =1-hydroxybenzotriazole hydrate hr = hour K₂CO₃ = potassium carbonate L =liter m = multiplet MeOH = methanol mg = milligram MgSO₄ = magnesiumsulfate mL = milliliter mm = millimeter mM = millimolar mmol = millimolmp = melting point N = normal NaCl = sodium chloride Na₂CO₃ = sodiumcarbonate NaHCO₃ sodium bicarbonate NaOEt = sodium ethoxide NaOH =sodium hydroxide NH₄Cl = ammonium chloride NMM = N-methylmorpholine Phe= L-phenylalanine Pro = L-proline psi = pounds per square inch PtO₂ =platinum oxide q = quartet quint. = quintet rt = room temperature s =singlet sat = saturated t = triplet t-BuOH = tert-butanol TFA =trifluoroacetic acid THF = tetrahydrofuran TLC or tlc = thin layerchromatography Ts = tosyl TsCl = tosyl chloride TsOH = tosylate μL =microliter

The following Methods may be used to prepare the compounds of thisinvention.

Method A 5-Trifluoromethylsulfonyloxy-2-methylpyridine PreparationProcedure

The procedure of Tilley et al. J. Org. Chem. 1998, 5, 3(2), 386-390, wasused to convert 5-hydroxy-2-methylpyridine into5-trifluoromethyl-sulfonyloxy-2-methylpyridine.

Method B Heteroaryl-aryl Preparation Procedure—Suzuki Coupling Procedure

A mixture of a heteroaryl halide or heteroaryl triflate (1.0 eq.), anarylboronic acid (1.1 eq.), CsCO₃ (1.5 eq.), Pd(PPh₃)₄ (0.03 eq.) Andanhydrous DMF was stirred under nitrogen at 100° C. for 48 h and thenthe DMF was evaporated. The residue was partitioned between ethylacetate and half-saturated aqueous sodium chloride and then the ethylacetate extracts were treated with magnesium sulfate, filtered andevaporated. The residue was purified by flash chromatography on silicagel using ethyl acetate/hexanes to give the heteroaryl-aryl product.

Method C 3-Heteroarylpyruvic Acid Ethyl Ester Preparation Procedure

To a stirred −78° C. solution of a methylheteroaryl compound (1 eq.) intetrahydrofuran was added dropwise 1.6 M n-butyllithium in hexanes (1.25eq.). The resulting dark red solution was warmed to 0° C. and thenre-cooled to −78° C. A solution of diethyl oxalate (1.25 eq.) in THF wasadded dropwise. The resulting mixture was warmed to 22° C., stirred for18 h diluted with aqueous HCl, and the extracted with methylenechloride. The methylene chloride extracts were treated with magnesiumsulfate, filtered, and evaporated. The residue was purified by flashchromatography on silica gel column using ethyl acetate/hexanes to givea 3-heteroarylpyruvic acid ethyl ester.

Method D 3-Heteroarylpyruvic Acid Ethyl Ester Oxime PreparationProcedure

The procedure of Negi et al. Synthesis, 1996, 8, 991, was used toconvert a 3-heteroarylpyruvic acid ethyl ester into a3-heteroarylpyruvic ethyl ester oxime.

Method E 3-Heteroarylalanine Acid Ethyl Ester Preparation Procedure

The procedure of Keck et al. Syn. Commun., 1979, 281-286 was used toconvert a 3-heteroarylpyruvic acid ethyl ester into aD,L-3-heteroaryvalanine ethyl ester.

Method F 3-Heteroaryldehydroalanine Methyl Ester Preparation Procedure

The reaction of N-benzyloxycarbonyldehydroalanine methyl ester with anaryl bromide according to the procedure of Carlstrom and Frejd. J. Org.Chem., 1991, 56, 1289-1293, was used to prepare a3-heteroaryldehydroalanine methyl ester.

Method G 3-Heteroarylalanine Methyl Ester Oxime Preparation Procedure

The procedure of Keck et al. Syn. Commun., 1979, 281-286 was used toconvert a 3-heteroaryldehydroalanine methyl ester into aD,L-3-heteroaryl-alanine methyl ester.

Method H Methyl Ester Preparation Procedure

Amino acid methyl esters can be prepared using the method of Brenner andHuber Helv. Chim. Acta 1953, 36, 1109.

Method I BOP Coupling Procedure

The desired dipeptide ester was prepared by the reaction of a carboxylicacid (1 equivalent) with the appropriate amino acid ester or amino acidester hydrochloride (1 equivalent),benzotriazol-1-yloxy-tris(dimethylamino)phos-phonium hexafluorophosphate[BOP] (2.0 equivalent), triethylamine (1.1 equivalent), and DMF. Thereaction mixture was stirred at room temperature overnight. The crudeproduct is purified flash chromatography to afford the dipeptide ester.

Method J Hydrogenation Procedure I

Hydrogenation was performed using 10% palladium on carbon (10% byweight) in methanol at 30 psi overnight. The mixture was filteredthrough a pad of Celite and the filtrate concentrated to yield thedesired compound.

Method K Hydrolysis Procedure I

To a chilled (0° C.) THF/H₂O solution (2:1, 5-10 mL) of the appropriateester was added LiOH (or NaOH) (0.95 equivalents). The temperature wasmaintained at 0° C. and the reaction was complete in 1-3 hours. Thereaction mixture was extracted with ethyl acetate and the aqueous phasewas lyophilized resulting in the desired carboxylate salt.

Method L Ester Hydrolysis Procedure II

To a chilled (0° C.) THF/H₂O solution (2:1, 5-10 mL) of the appropriateester was added LiOH (1.1 equivalents). The temperature was maintainedat 0° C. and the reaction was complete in 1-3 hours. The reactionmixture was concentrated and the residue was taken up into H₂O and thepH adjusted to 2-3 with aqueous HCl. The product was extracted withethyl acetate and the combined organic phase was washed with brine,dried over MgSO₄, filtered and concentrated to yield the desired acid.

Method M Ester Hydrolysis Procedure III

The appropriate ester was dissolved in dioxane/H₂O (1:1) and 0.9equivalents of 0.5 N NaOH was added. The reaction was stirred for 3-16hours and then concentrated. The resulting residue was dissolved in H₂Oand extracted with ethyl acetate. The aqueous phase was lyophilized toyield the desired carboxylate sodium salt.

Method N BOC Removal Procedure

Anhydrous hydrochloride (HCl) gas was bubbled through a methanolicsolution of the appropriate Boc-amino acid ester at 0° C. for 15 minutesand the reaction mixture was stirred for three hours. The solution wasconcentrated to a syrup and dissolved in Et₂O and reconcentrated. Thisprocedure was repeated and the resulting solid was placed under highvacuum overnight.

Method O tert-Butyl Ester Hydrolysis Procedure I

The tert-butyl ester was dissolved in CH₂Cl₂ and treated with TFA. Thereaction was complete in 1-3 hr at which time the reaction mixture wasconcentrated and the residue dissolved in H₂O and lyophilized to yieldthe desired acid.

Method P EDC Coupling Procedure I

To a CH₂Cl₂ solution (5-20 mL) of a carboxylic acid (1 equivalent), theappropriate amino acid ester hydrochloride (1 equivalent),N-methylmorpholine (1.1-2.2 equivalents) and 1-hydroxybenzotriazole (2equivalents) were mixed, placed in an ice bath and1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (1.1 equivalents) added.The reaction was allowed to rise to room temperature and stirredovernight. The reaction mixture was poured into H₂O and the organicphase was washed with sat. NaHCO₃, brine, dried (MgSO₄ or Na₂SO₄),filtered and concentrated. The crude product was purified by columnchromatography.

Method Q EDC Coupling Procedure II

To a DMF solution (5-20 mL) of a carboxylic acid (1 equivalent), theappropriated amino acid ester hydrochloride (1 equivalent), Et₃N (1.1equivalents) and 1-hydroxybenzotriazole (2 equivalents) were mixed,placed in an ice bath and 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide(1.1 equivalents) added. The reaction was allowed to rise to roomtemperature and stirred overnight. The reaction mixture was partitionedbetween EtOAc and H₂O and the organic phase washed with 0.2 N citricacid, H₂O, sat. NaHCO₃, brine, dried (MgSO₄ or Na₂SO₄), filtered andconcentrated. The crude product was purified by column chromatography orpreparative TLC.

Method R tert-Butyl Ester Hydrolysis Procedure II

The tert-butyl ester was dissolved in CH₂Cl₂ (5 mL) and treated with TFA(5 mL). The reaction was complete in 1-3 hours at which time thereaction mixture was concentrated and the residue dissolved in H₂O andconcentrated. The residue was redissolved in H₂O and lyophilized toyield the desired product.

Method S Carbamate Formation Procedure I

Into a reaction vial were combined 15.2 mmol, 1.0 eq. of the startinghydroxy compound (typically a tyrosine derivative) and 1.86 g (15.2mmol, 1.0 eq) DMAP. Methylene chloride (50 mL), triethylamine (2.12 mL,1.54 g, 15.2 mmol, 1.0 eq), and dimethylcarbamyl chloride (1.68 mL, 1.96g, 18.2 mmol, 1.2 eq) were then added. The vial was capped tightly, andthe reaction solution swirled to obtain a homogeneous solution. Thereaction solution was then heated to 40° C. After 48 h, TLC of theresulting colorless solution indicated complete conversion. The work-upof the reaction solution was as follows: 50 mL EtOAc and 50 mL hexaneswas added to the reaction mixture, and the resulting mixture was washedwith 0.5 M citric acid (3×50 mL), water (2×50 mL), 10% K₂CO₃ (2×50 mL),and sat. NaCl (1×50 mL); dried with MgSO₄, filtered and evaporated toafford the desired compound.

Method T Carbamate Formation Procedure II

Into a reaction vial were combined 84.34 mmol (1.0 eq) of the startinghydroxy compound (typically a tyrosine derivative) and 17.0 g (84.34mmol, 1.0 eq) 4-nitrophenyl chloroformate. Methylene chloride (700 mL)was added and the vial was capped with a septum. A nitrogen line wasattached and the vial was immersed in a 4:1 water/ethanol dry ice slurrywith stirring to cool to −15° C. Triethylamine (29.38 mL, 21.33 g,210.81 mmol, 2.5 eq) was added over five minutes with stirring and thestirring was continued at −10 to −15° C. for 1 h. N-Methyl piperazine(9.35 mL, 8.45 g, 84.34 mmol, 1.0 eq) was added over three minutes withstirring and stirring was continued overnight while warming to roomtemperature. The reaction mixture was diluted with 700 mL hexanes andthe resulting mixture was washed repeatedly with 10% KC₂CO₃, until noyellow color (from 4-nitrophenol) is observed in the aqueous layer. Themixture was then washed with sat. NaCl, dried over anhydrous MgSO₄,filtered and evaporated. The residue was dissolved in 500 mL of ethanoland evaporated to remove triethylamine. The residue was again dissolvedin 500 mL of ethanol and evaporated to remove triethylamine. The residuewas then dissolved in 400 mL of ethanol and 600 mL of water was addedwith stirring to precipitate a solid or oil. If an oil if formed, theoil is stirred vigorously to induce it to solidify. The solid is thenisolated by filtration. Dissolution, precipitation, and filtration arerepeated once and the resulting solid is rinsed with water to removetraces of yellow color. The solid is then subjected to high vacuum untilthe mass remains constant thereby affording the desired carbamyloxycompound.

Method U Preparation of 5-Iodo-4(3H)-pyrimidinone

The procedure of Sakamoto et. al. (Chem. Pharm. Bull. 1986, 34(7),2719-2724) was used to convert 4(3H)-pyrimidinone into5-iodo-4(3H)-pyrimidinone, which was of sufficient purity for conversionto 4-chloro-5-iodopyrimidine.

Method V Preparation of 4-Chloro-5-iodopyrimidine

5-Iodo-4(3H)-pyrimidinone (1 eq.) was suspended in toluene to which wasadded POCl₃ (2.0 eq.). The reaction mixture was heated to reflux for 3hours, and then cooled and concentrated. The residue was suspended inwater, adjusted to pH=7 by addition of 4N sodium hydroxide, andextracted with ethyl acetate. The organic extracts were washed withbrine, dried (MgSO₄), filtered and stripped to give a red oil. The crudeproduct was dissolved in methanol and silica gel was added. Followingconcentration, the coated silica gel was loaded onto a plug of silicagel and elution with ethyl acetate/hexanes yielded the title compound.

Method W Preparation of 4-N-Alkylamino-5-bromo-2-chloropyrimidine

A methanol solution of 5-bromo-2,4-dichloropyrimidine (1.0 eq), thealkylamine (1.05 eq, typicallyL-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butyl ester), andN,N-diisoproylethylamine (5.0 eq) was heated to 40° C. for 16 hours. Thereaction mixture was then concentrated, and the residue was taken up inethyl acetate. The organic portion was washed with 0.2 N citric acid,water, saturated NaHCO₃, brine, dried (MgSO₄), filtered andconcentrated. The crude material was purified by silica gelchromatography using ethyl acetate/hexanes to afford the desiredcompound.

Method X Preparation of 4-N-Alkylamino-5-bromo-2-N-alkylaminopyrimidine

An isopropanol solution of the 4-N-alkylamino-5-bromo-2-chloropyrimidine(1.0 eq) and an alkylamine (5.0 eq) was heated in sealed tube at 130° C.for 3-5 hours. The reaction mixture was then cooled and washed with 0.2N citric acid, water, saturated NaHCO₃, brine, dried (MgSO₄), filteredand concentrated. The crude material was purified by silica gelchromatography using ethyl acetatelhexanes to afford the desiredcompound.

Method Y 4-N-Alkylamino-5-bromo-2-N-alkylaminopyrimidine Suzuki CouplingProcedure

To an ethyleneglycol dimethyl ether solution oftetrakis(triphenylphosphine)palladium (0.04 eq) was added an4-N-alkylamino-5-bromo-2-N-alkylaminopyrimidine (1.0 eq.). Afterstirring for approximately ten minutes, the boronic acid or ester (1.2eq) and 2M Na₂CO₃ (2.0 eq) was added, and the reaction flask wasevacuated and then flushed with nitrogen gas. The reaction was heated atreflux for three to four hours. The reaction mixture was then cooled anddiluted with water and ethyl acetate, and the organic phase was washedwith 0.2 N citric acid, water, saturated NaHCO₃, brine, dried (MgSO₄),filtered and concentrated. The residue was purified by either silica gelcolumn or preparative thin layer chromatography using ethylacetate/hexanes to afford the desired product.

Method Z Preparation of Dimethyl 2-Alkylmalonate

To a suspension of sodium hydride 60% dispersion in mineral oil (1.1 eq)in anhydrous THF was added slowly with stirring dimethyl malonate (1.1eq), causing the evolution of gas. To the resulting solution was added abromoalkane, iodoalkane, or trifluoromethanesulfonyloxyalkane (1.0 eq),and the mixture was heated to 50° C. for 48 h, at which point TLCindicated consumption of the bromoalkane, iodoalkane, ortrifluoromethanesulfonyl-oxyalkane. The mixture was diluted with diethylether and washed with 70% saturated sodium chloride. The organicextracts were treated with anhydrous magnesium sulfate, filtered, andevaporated to afford a dimethyl 2-alkylmalonate of sufficient purity forimmediate conversion to a 5-alkyl-4,6-dihydroxypyrimidine.

Method AA Preparation of Diethyl 2-Alkylidenylmalonate

Procedure B (p. 2759) of Houve and Winberg (J. Org. Chem. 1980, 45(14),2754-2763) was employed to react diethyl malonate with a ketone or analdehyde to afford a diethyl 2-alkylidenylmalonate of sufficient purityfor immediate conversion to a diethyl 2-alkylmalonate.

Method BB Preparation of Diethyl 2-Alkylmalonate

A diethyl 2-alkylidenylmalonate and an equal mass 10% palladium oncarbon were suspended in ethanol. The mixture was shaken under 55 psihydrogen gas for 24 h, at which point TLC indicated consumption of thediethyl 2-alkylidenylmalonate. The mixture was filtered through Celiteand evaporated to afford a diethyl 2-alkylmalonate of sufficient purityfor immediate conversion to a 5-alkyl-4,6-dihydroxypyrimidine.

Method CC Preparation of 5-Alkyl-4,6-dihydroxypyrimidine

To a diethyl 2-alkylmalonate or a dimethyl 2-alkylmalonate (1.0 eq) wasadded formamidine acetate (1.0 eq) and 25% sodium methoxide in methanol(3.3 eq). The resulting slurry was stirred vigorously and heated to 60°C. for 4 h, and then allowed to cool. The slurry was diluted with water,and acidified to pH=2 by addition of HCl. The resulting precipitate wascollected by filtration, washed with water, and dried under vacuum, toafford a 5-alkyl-4,6-dihydroxypyrimidine of sufficient purity forimmediate conversion to a 5-alkyl-4,6-dichloropyrimidine.

Method DD Preparation of 5-Alkoxy-4-hydroxypyrimidine

The method (p. 308) of Anderson et al. (Org. Proc. Res. Devel. 1997, 1,300-310) was employed to react a methyl alkoxyacetate, sodium methoxide,ethyl formate, and formamidine acetate to afford a5-alkoxy-4-hydroxypyrimidine of sufficient purity for immediateconversion to a 5-alkoxy-4-chloropyrimidine.

Method EE Preparation of 5-Alkyl-4,6-dichloropyrimidine or5-Alkoxy-4-chloropyrimidine

To a 5-alkyl-4,6-dihydroxypyrimidine or a 5-alkoxy-4-hydroxypyrimidine(1.0 eq) were added phosphorus oxychloride (15.0 eq) andN,N-dimethylaniline (1.0 eq), and the mixture was heated to 100° C. for3 h, and then allowed to cool. The resulting solution was poured ontoice, and the mixture was extracted with dichloromethane. The organicextracts were treated with anhydrous magnesium sulfate, filtered, andevaporated to afford a 5-alkyl-4,6-dichloropyrimidine or a5-alkoxy-4-chloropyrimidine of sufficient purity for immediateconversion to a 5-alkyl-4-N-alkylamino-6-chloropyrimidine or a5-alkoxy-4-N-alkylaminopyrimidine.

Method FF Preparation of 5-Alkyl-4-N-alkylamino-6-chloropyrimidine or5-Alkoxy-4-N-alkylaminopyrimidine

To a solution of a 5-alkyl-4,6-dichloropyrimidine or a5-alkoxy-4-chloropyrimidine (1.0 eq) in ethanol were added an alkylamine (1.2 eq, typically L-4-(N,N-dimethylcarbamyloxy)-phenylalaninetert-butyl ester) and diisopropylethylamine (2.0 eq). The mixture wassealed in a pressure tube and heated to 120° C. for 48 h, at which pointTLC indicated consumption of the 5-alkyl-4,6-dichloropyrimidine or the5-alkoxy-4-chloropyrimidine. The mixture was evaporated, and the residuewas partitioned between ethyl acetate and pH=4.5 citrate buffer. Theorganic extracts were washed with saturated sodium chloride, treatedwith anhydrous magnesium sulfate, filtered, and evaporated. The residuewas purified by chromatography on silica gel using ethyl acetate andhexanes to afford a pure 5-alkyl-4-N-alkylamino-6-chloropyrimidine or5-alkoxy-4-N-alkylaminopyrimidine.

Method GG Preparation of 5-Alkyl-4-N-alkylaminopyrimidine (Procedure I)

A suspension of 5-alkyl-4-N-alkylamino-6-chloropyrimidine (1.0 eq), andan equal mass 10% palladium on carbon, and sodium bicarbonate (5.0 eq)in methanol was shaken under 55 psi hydrogen gas for 16 h, at whichpoint TLC indicated consumption of the5-alkyl-4-N-alkylamino-6-chloropyrimidine. The mixture was filteredthrough Celite and evaporated to give a residue, which was partitionedbetween ethyl acetate and 70% saturated sodium chloride. The organicextracts were treated with anhydrous magnesium sulfate, filtered, andevaporated. The residue was purified by chromatography on silica gelusing ethyl acetate and hexanes to afford a pure5-alkyl-4-N-alkylaminopyrimidine.

Method HH Preparation of 5-Alkyl-4-N-alkylaminopyrimidine (Procedure II)

A suspension of 5-alkyl-4-N-alkylamino-6-chloropyrimidine (1.0 eq),sodium acetate (10.0 eq), and zinc powder (20.0 eq) in a 9:1 mixture ofacetic acid and water was stirred vigorously at 40° C. for 72 h, atwhich point TLC indicated partial consumption of the5-alkyl-4-N-alkylamino-6-chloropyrimidine. The supernatant solution wasdecanted from remaining zinc and evaporated. The residue was partitionedbetween ethyl acetate and saturated sodium bicarbonate, and the organicextracts were treated with anhydrous magnesium sulfate, filtered, andevaporated. The residue was purified by chromatography on silica gelusing ethyl acetate and hexanes to afford a pure5-alkyl-4-N-alkylaminopyrimidine.

Method II Preparation of 2,4-Dichloro-5-nitropyrimidine

5-Nitrouracil was treated with phosphorus oxychloride andN,N-dimethylaniline, according to the procedure of Whittaker (J. Chem.Soc. 1951, 1565), to give 2,4-dichloro-5-nitropyrimidine as an orangeoil, which was used without distillation immediately in the next step.

Method JJ Preparation of Diethyl 2-(N,N-Dialkylamino)malonate

The appropriate amine (1.0 eq) was added to a 0° C. solution of diethylbromomalonate (1.0 eq) and N,N-diisopropylethyl amine (1.1 eq) inethanol. The mixture was stirred and allowed to warm room temperature.After 16 hours, the reaction mixture was concentrated and the residuewas suspended in ethyl acetate and sat. NaHCO₃. The organic portion waswashed with sat NaHCO₃, brine, dried (MgSO₄) filtered and concentratedto yield the diethyl 2-(N,N-dialkylamino)malonate, of sufficient purityfor immediate conversion to a5-(N,N-dialkylamino)-4,6-dihydroxypyrimidine.

Method KK Preparation of 5-(N,N-Dialkylamino)-4,6-dihydroxypyrimidine

A suspension of a diethyl 2-N,N-dialkylamino)malonate (1.0 eq),formamidine acetate (1.10 eq.) and 25% sodium methoxide in methanol (3.3eq) was heated to 65° C. for 3.5 hours. The reaction mixture was cooledand diluted with water. The mixture was acidified to pH=4.5 by additionof dilute HCl. The resulting precipitate was collected by filtration,washed with water, and dried under vacuum to afford a5-(N,N-dialkylamino)-4,6-dihydroxy-pyrimidine of sufficient purity forimmediate conversion to a 5-(N,N-dialkylamino)-4,6-dichloropyrimidine.Alternatively, the acidified solution was evaporated to give a solidresidue, which was extracted with boiling ethanol. The ethanol extractswere filtered and concentrated to give a residue, which wasrecrystallized from isopropyl alcohol to afford a5-(N,N-dialkylamino)-4,6-dihydroxypyrimidine of sufficient purity forimmediate conversion to a 5-(N,N-dialkylamino)-4,6-dichloropyrimidine.

Method LL Preparation of 5-(N,N-Dialkylamino)-4,6-dichloropyrimidine

A 5-(N,N-dialkylamino)-4,6-dihydroxypyrimidine (1.0 eq) was suspended inPOCl₃ (15.0 eq), and the mixture was heated to reflux for 16 hours. Thenthe mixture was cooled and carefully poured into a suspension of ethylether and aqueous K₂CO₃. The organic portion was washed with brine,dried (MgSO₄), filtered and concentrated to yield a5-(N,N-dialkylamino)-4,6-dichloro-pyrimidine of sufficient purity forimmediate reaction with alkylamines.

Method MM Preparation of4-(N-Alkylamino)-5-(N,N-dialkylamino)-6-chloropyrimidine

A 5-(N,N-dialkylamino)-4,6-dichloropyrimidine (1.0 eq),L-4-(N,N-dimethylcarbamyloxy)phenylalanine tert-butyl ester (1.5 eq) andN,N-diisopropyl ethylamine (1.5 eq) were dissolved in ethanol and heatedto 120° C. in a sealed tube for 72 h. The cooled reaction mixture wasconcentrated, and the residue dissolved in ethyl acetate. The ethylacetate solution was washed with 0.2 N citric acid, water, saturatedNaHCO₃, brine, dried (MgSO₄), filtered and concentrated. The residue waspurified by silica gel chromatography using ethyl acetate/hexanes toafford the 4-(N-alkylamino)-5-(N,N-dialkylamino)-6-chloropyrimidine.

Method NN Preparation of 4-(N-Alkylamino)-5-(N,N-dialkylamino)pyrimidine

A 4-(N-Alkylamino)-5-(N,N-dialkylamino)-6-chloropyrimidine (1.0 eq), anequal mass of 10% palladium on carbon and NaHCO₃ (5.0 eq) were suspendedin methanol. The reaction mixture was hydrogenated at 45 psi hydrogenfor 16 hours and then filtered through a pad of Celite. The filtrate wasconcentrated, and the residue was dissolved in ethyl acetate. The ethylacetate solution was washed with water, brine, dried (MgSO₄), filteredand concentrated to yield an oil. The oil was purified by columnchromatorgraphy on silica gel using ethyl actate and hexanes to afford apure 4-(N-alkylamino)-5-(N,N-dialkylamino)pyrimidine.

Method OO Bromopyrimidine Debromination Procedure

The bromopyrimidine was dissolved in isopropyl alcohol to which wasadded 10% palladium on carbon. The reaction was hydrogenated at 45 psihydrogen. Filtration and concentration of the filtrate yielded thedesired dehalogenated pyrimidine.

Method PP Preparation of 2-Isopropropoxypyrimidine

A 2-chloropyrimidine was dissolved in isopropyl alcohol to which wasadded diisopropylamine. The reaction was heated in a sealed tube for tendays at 130° C. The cooled reaction mixture was concentrated, and theproduct purified via silica gel column chromatography to yield the2-isopropoxypyrimidine.

Method QQ Preparation of 2-Amino-3-Chloropyrazine

A mixture of 2,3-dichloropyrazine (Lancaster) and ammonium hydroxide washeated in a sealed tube at 100° C. for 24 h resulting in a whiteprecipitate. The precipitate was collected by filtration and dried undervacuum to afford 2-amino-3-chloropyrazine of sufficient purity forimmediate conversion to 2-chloro-3-nitropyrazine.

Method RR Preparation of 2-Chloro-3-Nitropyrazine

The method (p. 1638) of Hartnan et al. (J. Med. Chem. 1984, 27(12),1634-1639) was employed to convert 2-amino-3-chloropyrazine into2-chloro-3-nitropyrazine of sufficient purity for immediate use.

Method SS Preparation of 4-Alkylamino-2-dialkylamino-5-nitropyrimidine

A solution of 1.0 eq 4-alkylamino-2-chloro-5-nitropyrimidine and 5.0 eqdialkylamine in THF was allowed to stand for 16 h. The mixture wasdiluted with ethyl acetate and then washed with pH=4.5 citrate bufferand saturated sodium chloride. The organic extracts were treated withanhydrous magnesium sulfate, filtered, and evaporated to give a residue,which was purified by chromatography on silica gel using ethyl acetateand hexanes.

Method TT Preparation of 5-Hydroxypyridin-2-ylalanine

The methods of Schow et al. (J. Org. Chem. 1994 59:22, 6850-6852), Ye etal (J. Org. Chem. 1995 60:8, 2640-2641) or Myers et al. (J. Org. Chem.1996 61:22, 813-815) may be used to prepare the title compound.

Method UU Preparation of 2-Hydroxypyridin-5-ylalanine

The methods of Yamano et al. (Tetrahedron. 1992 48:8, 1457-1464) orAndrews et al. (J. Chem. Soc. Perkin Trans. 1995 11, 1335-1340) may beused to prepare the title compound.

Example 1 Synthesis ofN-(5-(2,2,2-trifluoroethyl)pyrimidin-4-yl)-D,L-3-(5-(2,5-dimethoxyphenyl)-pyridin-2-yl)alanine

5-Hydroxy-2-methylpyridine was converted via method A to give5-trifluoromethylsulfonyloxy-2-methylpyridine. This compound and2,6-dimethoxyphenylboronic acid were coupled via method B to give5-(2,6-dimethoxyphenyl)-2-methylpyridine which was then converted viasequential application of methods C,D and E to giveD,L-(5-(2,6-dimethoxy-phenyl)pyridin-2-yl)alanine ethyl ester.

2,2,2-Trifluoroethyl (prepared accordingly to Gassman, et al. J. Org.Chem., 1984, 49(12), 2258-2273) was converted via sequential applicationof methods Z, CC, and EE to give4,6-dichloro-5-(2,2,2-trifluoroethyl)pyrimidine.D,L-(5-(2,6-dimethoxyphenyl)pyridine-2-yl)alanine ethyl ester and4,6-dichloro-5-(2,2,2-trifluoroethyl)pyrimidine were coupled via methodFF and the product was transformed via sequential application of methodsGG and L to give the title compound.

Example 2 Synthesis ofN-(5-(2,2,2-trifluoroethyl)pyrimidin-4-yl)-D,L-3-(5-(2-methoxyphenyl)-pyridin-2-yl)alanine

5-Hydroxy-2-methylpyridine was converted via method A to give5-trifluoromethylsulfonyloxy-2-methylpyridine. This compound and2-methoxyphenylboronic acid were coupled via method B to give5-(2-methoxyphenyl)-2-methylpyridine which was then converted viasequential application of methods C,D and E to giveD,L-(5-(2-methoxyphenyl)pyridin-2-yl)alanine ethyl ester.

2,2,2-Trifluoroethyl (prepared accordingly to Gassman, et al. J. Org.Chem., 1984, 49(12), 2258-2273) was converted via sequential applicationof methods Z, CC, and EE to give4,6-dichloro-5-(2,2,2-trifluoroethyl)pyrimidine.D,L-(5-(2-methoxyphenyl)pyridine-2-yl)alanine ethyl ester and4,6-dichloro-5-(2,2,2-trifluoroethyl)pyrimidine were coupled via methodFF and the product was transformed via sequential application of methodsGG and L to give the title compound.

Example 3 Synthesis ofN-(5-(2,2,2-trifluoroethyl)pyrimidin-4-yl)-D,L-3-(5-(N,N-dimethylaminocarbonyloxy)-pyridin-2-yl)alanine

5-Hydroxypyridin-2-ylalanine, which can be prepared as per Example TT,is N and C protected via conventional methods to provide for ethyl N-Boc5-hydroxypyridin-2-ylalanine. This compound is converted either bymethods S or T to provide for ethyl N-Boc5-(N,N-dimethylaminocarbonyloxypyridin-2-ylalanine. Conventionaldeprotection provides for 5-(N,Ndimethylaminocarbonyloxypyridin-2-ylalanine.

Coupling of the amino of the5-(N,N-dimethylaminocarbonyloxypyridin-2-ylalanine with4,6-dichloro-5-(2,2,2-trifluoroethyl)pyrimidine as per Example 2 abovewill provide the title compound.

Example 4 Synthesis ofN-(5-(2,2,2-trifluoroethyl)pyrimidin-4-yl)-D,L-3-(2-(N,N-dimethylaminocarbonyloxy)-pyridin-5-yl)alanine

2-Hydroxypyridin-5-ylalanine, which can be prepared as per Example UU,is N and C protected via conventional methods to provide for ethyl N-Boc2-hydroxypyridin-5-ylalanine. This compound is converted either bymethods S or T to provide for ethyl N-Boc2-(N,N-dimethylaminocarbonyloxypyridin-5-ylalanine. Conventionaldeprotection provides for2-(N,N-dimethylaminocarbonyloxypyridin-5-ylalanine.

Coupling of the amino of the2-(N,N-dimethylaminocarbonyloxypyridin-5-ylalanine with4,6-dichloro-5-(2,2,2-trifluoroethyl)pyrimidine as per Example 2 abovewill provide the title compound.

Biological Examples Example A In vitro Assay For Determining Binding ofCandidate Compounds to VLA-4

An in vitro assay was used to assess binding of candidate compounds toα₄β₁ integrin. Compounds which bind in this assay can be used to assessVCAM-1 levels in biological samples by conventional assays (e.g.,competitive assays). This assay is sensitive to IC₅₀ values as low asabout 1 nM.

The activity of α₄β₁ integrin was measured by the interaction of solubleVCAM-1 with Jurkat cells (e.g., American Type Culture Collection Nos.TIB 152, TIB 153, and CRL 8163), a human T-cell line which expresseshigh levels of α₄β₁ integrin. VCAM-1 interacts with the cell surface inan α₄β₁ integrin-dependent fashion (Yednock, et al. J. Biol. Chem.,1995, 270:28740).

Recombinant soluble VCAM-1 was expressed as a chimeric fusion proteincontaining the seven extracellular domains of VCAM-1 on the N-terminusand the human IgG₁ heavy chain constant region on the C-terminus. TheVCAM-1 fusion protein was made and purified by the manner described byYednock, supra.

Jurkat cells were grown in RPMI 1640 supplemented with 10% fetal bovineserum, penicillin, streptomycin and glutamine as described by Yednock,supra.

Jurkat cells were incubated with 1.5 mM MnCl₂ and 5 μg/mL 15/7 antibodyfor 30 minutes on ice. Mn⁺² activates the receptor to enhance ligandbinding, and 15/7 is a monoclonal antibody that recognizes anactivated/ligand occupied conformation of α₄β₁ integrin and locks themolecule into this conformation thereby stabilizing the VCAM-1/α₄β₁integrin interaction. Yednock, et al., supra. Antibodies similar to the15/7 antibody have been prepared by other investigators (Luque, et al,1996, J. Biol. Chem. 271:11067) and may be used in this assay.

Cells were then incubated for 30 minutes at room temperature withcandidate compounds, in various concentrations ranging from 66 μM to0.01 μM using a standard 5-point serial dilution. 15 μL solublerecombinant VCAM-1 fusion protein was then added to Jurkat cells andincubated for 30 minutes on ice. (Yednock et al., supra.).

Cells were then washed two times and resuspended in PE-conjugated goatF(ab′)₂ anti-mouse IgG Fc (Immunotech, Westbrook, Me.) at 1:200 andincubated on ice, in the dark, for 30 minutes. Cells were washed twiceand analyzed with a standard fluorescence activated cell sorter (“FACS”)analysis as described in Yednock, et al., supra.

Compounds having an IC₅₀ of less than about 15 μM possess bindingaffinity to α₄β₁.

When tested in this assay, each of the compound prepared in the aboveexamples has or is expected to have an IC₅₀ of 15 μM or less (or isexpected to be active in vivo).

Example B In Vitro Saturation Assay for Determining Binding of CandidateCompounds to α₄β₁

The following describes an in vitro assay to determine the plasma levelsneeded for a compound to be active in the Experimental AutoimmuneEncephalomyelitis (“EAE”) model, described in the next example, or inother in vivo models.

Log-growth Jurkat cells are washed and resuspended in normal animalplasma containing 20 μg/ml of the 15/7 antibody (described in the aboveexample).

The Jurkat cells are diluted two-fold into either normal plasma samplescontaining known candidate compound amounts in various concentrationsranging from 66 μM to 0.01 μM, using a standard 12 point serial dilutionfor a standard curve, or into plasma samples obtained from theperipheral blood of candidate compound-treated animals.

Cells are then incubated for 30 minutes at room temperature, washedtwice with phosphate-buffered saline (“PBS”) containing 2% fetal bovineserum and 1 mM each of calcium chloride and magnesium chloride (assaymedium) to remove unbound 15/7 antibody.

The cells are then exposed to phycoerythrin-conjugated goat F(ab′)₂anti-mouse IgG Fc (Immunotech, Westbrook, Me.), which has been adsorbedfor any non-specific cross-reactivity by co-incubation with 5% serumfrom the animal species being studied, at 1:200 and incubated in thedark at 4° C. for 30 minutes.

Cells are washed twice with assay medium and resuspended in the same.They are then analyzed with a standard fluorescence activated cellsorter (“FACS”) analysis as described in Yednock et al. J. Biol. Chem.,1995, 270:28740.

The data is then graphed as fluorescence versus dose, e.g., in a normaldose-response fashion. The dose levels that result in the upper plateauof the curve represent the levels needed to obtain efficacy in an invivo model.

This assay may also be used to determine the plasma levels needed tosaturate the binding sites of other integrins, such as the α₉β₁integrin, which is the integrin most closely related α₄β₁ (Palmer et al,1993, J. Cell Bio., 123:1289). Such binding is predictive of in vivoutility for inflammatory conditions mediated by α₉β₁ integrin, includingby way of example, airway hyper-responsiveness and occlusion that occurswith chronic asthma, smooth muscle cell proliferation inatherosclerosis, vascular occlusion following angioplasty, fibrosis andglomerular scarring as a result of renal disease, aortic stenosis,hypertrophy of synovial membranes in rheumatoid arthritis, andinflammation and scarring that occur with the progression of ulcerativecolitis and Crohn's disease.

Accordingly, the above-described assay may be performed with a humancolon carcinoma cell line, SW 480 (ATTC #CCL228) transfected with cDNAencoding α₉ integrin (Yokosaki et al., 1994, J. Biol. Chem., 269:26691),in place of the Jurkat cells, to measure the binding of the α₉β₁integrin. As a control, SW 480 cells which express other α and β₁subunits may be used.

Accordingly, another aspect of this invention is directed to a methodfor treating a disease in a mammalian patient, which disease is mediatedby α₉β₁, and which method comprises administering to said patient atherapeutically effective amount of a compound of this invention. Suchcompounds are preferably administered in a pharmaceutical compositiondescribed herein above. Effective daily dosing will depend upon the age,weight, condition of the patient which factors can be readilyascertained by the attending clinician. However, in a preferredembodiment, the compounds are administered from about 20 to 500 μg/kgper day.

Example C In vivo Evaluation

The standard multiple sclerosis model, Experimental Autoimmune (orAllergic) Encephalomyelitis (“EAE”), was used to determine the effect ofcandidate compounds to reduce motor impairment in rats or guinea pigs.Reduction in motor impairment is based on blocking adhesion betweenleukocytes and the endothelium and correlates with anti-inflammatoryactivity in the candidate compound. This model has been previouslydescribed by Keszthelyi et al., Neurology, 1996, 47:1053-1059, andmeasures the delay of onset of disease.

Brains and spinal cords of adult Hartley guinea pigs were homogenized inan equal volume of phosphate-buffered saline. An equal volume ofFreund's complete adjuvant (100 mg mycobacterium tuberculosis plus 10 mlFreund's incomplete adjuvant) was added to the homogenate. The mixturewas emulsified by circulating it repeatedly through a 20 ml syringe witha peristaltic pump for about 20 minutes.

Female Lewis rats (2-3 months old, 170-220 g) or Hartley guinea pigs (20day old, 180-200 g) were anesthetized with isoflurane and threeinjections of the emulsion, 0.1 ml each, were made in each flank. Motorimpairment onset is seen in approximately 9 days.

Candidate compound treatment began on Day 8, just before onset ofsymptoms. Compounds were administered subcutaneously (“SC”), orally(“PO”) or intraperitoneally (“IP”). Doses were given in a range of 10mg/kg to 200 mg/kg, bid, for five days, with typical dosing of 10 to 100mg/kg SC, 10 to 50 mg/kg PO, and 10 to 100 mg/kg IP.

Antibody GG5/3 against α₄β₁ integrin (Keszthelyi et al., Neurology,1996, 47:1053-1059), which delays the onset of symptoms, was used as apositive control and was injected subcutaneously at 3 mg/kg on Day 8 and11.

Body weight and motor impairment were measured daily. Motor impairmentwas rated with the following clinical score:

0 no change 1 tail weakness or paralysis 2 hindlimb weakness 3 hindlimbparalysis 4 moribund or dead

A candidate compound was considered active if it delayed the onset ofsymptoms, e.g., produced clinical scores no greater than 2 or slowedbody weight loss as compared to the control.

Example D Asthma Model

Inflammatory conditions mediated by α₄β₁ integrin include, for example,airway hyper-responsiveness and occlusion that occurs with chronicasthma. The following describes an asthma model which can be used tostudy the in vivo effects of the compounds of this invention for use intreating asthma.

Following the procedures described by Abraham et al, J. Clin. Invest,93:776-787 (1994) and Abraham et al, Am J. Respir Crit Care Med,156:696-703 (1997), both of which are incorporated by reference in theirentirety. Compounds of this invention are formulated into an aerosol andadministered to sheep which are hypersensitive to Ascaris suum antigen.Compounds which decrease the early antigen-induced bronchial responseand/or block the late-phase airway response, e.g., have a protectiveeffect against antigen-induced late responses and airwayhyper-responsiveness (“AHR”), are considered to be active in this model.

Allergic sheep which are shown to develop both early and late bronchialresponses to inhaled Ascaris suum antigen are used to study the airwayeffects of the candidate compounds. Following topical anesthesia of thenasal passages with 2% lidocaine, a balloon catheter is advanced throughone nostril into the lower esophagus. The animals are then intubatedwith a cuffed endotracheal tube through the other nostril with aflexible fiberoptic bronchoscope as a guide.

Pleural pressure is estimated according to Abraham (1994). Aerosols (seeformulation below) are generated using a disposable medical nebulizerthat provides an aerosol with a mass median aerodynamic diameter of 3.2μm as determined with an Andersen cascade impactor. The nebulizer isconnected to a dosimeter system consisting of a solenoid valve and asource of compressed air (20 psi). The output of the nebulizer isdirected into a plastic T-piece, one end of which is connected to theinspiratory port of a piston respirator. The solenoid valve is activatedfor 1 second at the beginning of the inspiratory cycle of therespirator. Aerosols are delivered at V_(T) of 500 ml and a rate of 20breaths/minute. A 0.5% sodium bicarbonate solution only is used as acontrol.

To assess bronchial responsiveness, cumulative concentration-responsecurves to carbachol can be generated according to Abraham (1994).Bronchial biopsies can be taken prior to and following the initiation oftreatment and 24 hours after antigen challenge. Bronchial biopsies canbe preformed according to Abraham (1994).

An in vitro adhesion study of alveolar macrophages can also be performedaccording to Abraham (1994), and a percentage of adherent cells iscalculated.

Aerosol Formulation

A solution of the candidate compound in 0.5% sodium bicarbonate/saline(w/v) at a concentration of 30.0 mg/mL is prepared using the followingprocedure:

A. Preparation of 0.5% Sodium Bicarbonate/Saline Stock Solution: 100.0mL.

Ingredient Gram/100.0 mL Final Concentration Sodium Bicarbonate 0.5 g0.5% Saline q.s. ad 100.0 mL q.s. ad 100%

Procedure:

1. Add 0.5 g sodium bicarbonate into a 100 mL volumetric flask.

2. Add approximately 90.0 mL saline and sonicate until dissolved.

3. Q.S. to 100.0 mL with saline and mix thoroughly.

B. Preparation of 30.0 mg/mL Candidate Compound: 10.0 mL

Ingredient Gram/10.0 mL Final Concentration Candidate Compound 0.300 g30.0 mg/mL 0.5% Sodium q.s. ad 10.0 mL q.s ad 100% Bicarbonate/SalineStock Solution

Procedure:

1. Add 0.300 g of the candidate compound into a 10.0 mL volumetricflask.

2. Add approximately 9.7 mL of 0.5% sodium bicarbonate/saline stocksolution.

3. Sonicate until the candidate compound is completely dissolved.

4. Q.S. to 10.0 mL with 0.5% sodium bicarbonate/saline stock solutionand mix thoroughly.

Using a conventional oral formulation, compounds of this invention wouldbe active in this model.

Example E Allograft Model

Allograft rejection, associated with infiltration of inflammatory cells,is the leading obstacle to long-term allograft survival. Cell surfaceadhesion molecules facilitate alloantigen recognition in vitro and maybe critical for lymphocyte traffic in vivo. The following describes amodel which can be used to study the in vivo effects of the compounds ofthis invention in the control of allograft rejection.

The following procedures are described in Coito et al., Transplantation(1998) 65(6):699-706 and in Korom et al., Transplantation (1998)65(6):854-859, both of which are incorporated by reference in theirentirety.

Following the procedures described in Coito and Korom, male adult ratsweighing approximately 200-250 g are used in this model. Lewis rats areused as the recipients of cardiac allografts from Lewis X Brown Norwayrats. Hearts are transplanted into the abdominal great vessels usingstandard microvascular techniques.

A candidate compound is administered to the transplant recipient in asuitable pharmaceutical carrier for a 7-day course of treatment startingthe day of the engraftment. Doses range from 0.3 to 30 mg/kg/day.Control recipients receive the pharmaceutical carrier only. The rats areeuthanized and their cardiac allografts are analyzed as described inCoito and Korom.

Using conventional formulations, compounds of this invention would beactive in this model.

1. A compound of Formula (I):

wherein: A is an aryl, heteroaryl, cycloalkyl, or heterocyclic groupwherein said aryl, heteroaryl, cycloalkyl, or heterocyclic group isoptionally substituted, on any ring atom capable of substitution, with1-3 substituents selected from the group consisting of alkyl,substituted alkyl, alkoxy, substituted alkoxy, acyl, acylamino,thiocarbonylamino, acyloxy, amino, substituted amino, amidino, alkylamidino, thioamidino, aminoacyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, aryl, substituted aryl,aryloxy, substituted aryloxy, aryloxyaryl, substituted aryloxyaryl,cyano, halogen, hydroxyl, nitro, oxo, carboxyl, cycloalkyl, substitutedcycloalkyl, guanidino, guanidinosulfone, thiol, thioalkyl, substitutedthioalkyl, thioaryl, substituted thioaryl, thiocycloalkyl, substitutedthiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,thioheterocyclic, substituted thioheterocyclic, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —OS(O)₂-alkyl, —OS(O)₂-substituted alkyl,—OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where each R isindependently hydrogen or alkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substitutedalkyl, —NRS(O)₂-aryl, —NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl,—NRS(O)₂-substituted heteroaryl, —NRS(O)₂-heterocyclic,—NRS(O)₂-substituted heterocyclic, —NRS(O)₂—NR-alkyl,NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl, —NRS(O)₂—NR-substitutedaryl, —NRS(O)₂—NR-heteroaryl, —NRS(O)₂—NR-substituted heteroaryl,—NRS(O)₂—NR-heterocyclic, —NRS(O)₂—NR-substituted heterocyclic where Ris hydrogen or alkyl, —N[S(O)₂—R′]₂ and —N[S(O)₂—NR′]₂ where each R′ isindependently selected from the group consisting of alkyl, substitutedalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic; HetAr is a nitrogencontaining heteroaryl or a nitrogen containing substituted heteroarylgroup; Alk is an alkylene group of 1 to 4 carbons; m is 0 or 1; R¹ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic and substitutedheterocyclic; X is selected from the group consisting of hydroxyl,alkoxy, substituted alkoxy, alkenoxy, substituted alkenoxy, cycloalkoxy,substituted cycloalkoxy, cycloalkenoxy, substituted cycloalkenoxy,aryloxy, substituted aryloxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy and —NR″R″ where each R″ isindependently selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, substitutedcycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic; and enantiomers, diasteromersand pharmaceutically acceptable salts thereof; and further wherein thecompound of Formula (I) has a binding affinity to VLA-4 as expressed byan IC₅₀ of about 15μM or less.
 2. The compound of claim 1 wherein HetAris a nitrogen containing substituted heteroaryl group.
 3. The compoundof claim 1 wherein HetAr is a nitrogen containing heteroaryl group thatis substituted with a substituent selected from the group consisting ofacyl, acylamino, acyloxy, aminoacyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, oxycarbonylamino,oxythiocarbonylamino, thioamidino, thiocarbonylamino,aminosulfonylamino, aminosulfonyloxy, aminosulfonyl, oxysulfonylaminooxysulfonyl, aryl and substituted aryl.
 4. The compound of claim 1wherein HetAr is a nitrogen containing heteroaryl group is substitutedwith a group of formula —O-Z-NR¹¹R^(11′) or —O-Z-R¹² wherein R¹¹ andR^(11′) are independently selected from the group consisting ofhydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heterocyclic, substitutedheterocyclic, and where R¹¹ and R^(11′) are joined to form a heterocycleor a substituted heterocycle, R¹² is selected from the group consistingof heterocycle and substituted heterocycle, and Z is selected from thegroup consisting of —C(O)— and —SO₂—.
 5. The compound of claim 4 whereinthe nitrogen containing heteroaryl group is substituted with a group offormula —OC(O)NR¹¹R^(11′) wherein R¹¹ and R^(11′) are independentlyselected from the group consisting of alkyl or R¹¹ and R^(11′) arejoined to form a heterocycle or a substituted heterocycle.
 6. Thecompound of claim 5 wherein the nitrogen containing heteroaryl group issubstituted with —OC(O)N(CH₃)₂.
 7. The compound of claim 1 wherein HetAris a nitrogen containing heteroaryl group is subsituted with an aryl orsubstituted aryl group.
 8. The compound of claim 1 wherein A is aheteroaryl group which is optionally substituted with 1 to 3substituents selected from the group consisting of alkyl, substitutedalkyl, alkoxy, substituted alkoxy, amino, substituted amino, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic and halogen.
 9. Thecompound of claim 8 wherein A is 1-oxo-1,2,5-thiadiazole,1,1-dioxo-1,2,5-thiadiazole, pyridazine, pyrimidine or pyrazine ringwhich is optionally substituted with 1 to 3 substituents selected fromthe group consisting of alkyl, substituted alkyl, alkoxy, substitutedalkoxy, amino, substituted amino, cycloalkyl, substituted cycloalkyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic and halogen.
 10. The compound ofclaims 1 to 9 wherein R¹ is hydrogen, and X is hydroxyl.
 11. Thecompound of claim 1 wherein the compound has formula IIa, IIb, IIc, IId,or IIe:

wherein: HetAr is a nitrogen containing heteroaryl group substitutedwith a substituent selected from the group consisting of acyl,acylamino, acyloxy, aminoacyl, aminocarbonylamino,aminothiocarbonylamino, aminocarbonyloxy, oxycarbonylamino,oxythiocarbonylamino, tioamidino, thiocarbonylamino, aminosulfonylamino,aminosulfonyloxy, aminosulfonyl, oxysulfonylamino, aryl, substitutedaryl, and oxysulfonyl; R⁵ is selected from the group consisting ofalkyl, substituted alkyl, alkenyl, substituted alkenyl, aryl,substituted aryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, heterocyclic, substituted heterocylic,heteroaryl and substituted heteroaryl; R⁶ is selected from the groupconsisting of hydrogen, alkyl, substituted alkyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl,heterocyclic, substituted heterocyclic, aryl, substituted aryl,heteroaryl, substituted heteroaryl, and —SO₂R¹⁰ where R¹⁰ is selectedfrom the group consisting of alkyl, substituted alkyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl,heterocyclic, substituted heterocyclic, aryl, substituted aryl,heteroaryl, substituted heteroaryl; R⁷ and R⁸ are independently selectedfrom the group consisting of hydrogen, alkyl, substituted alkyl,cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, substituted heterocyclic andhalogen; R¹⁶ and R¹⁷ are independently selected from the groupconsisting of hydrogen, alkyl, substituted alkyl, alkoxy, substitutedalkoxy, amino, substituted amino, cycloalkyl, substituted cycloalkyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic and halogen; and R¹⁸ is selectedfrom the group consisting of alkyl, substituted alkyl, alkoxy,substituted alkoxy, amino, substituted amino, cycloalkyl, substitutedcycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic; R²⁰ is selected from thegroup consisting of hydrogen, alkyl, substituted alkyl, alkoxy,substituted alkoxy, cycloalkyl, substituted cycloalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic,substituted heterocyclic and halogen; R²¹ is selected from the groupconsisting of alkyl, substituted alkyl, alkoxy, substituted alkoxy,amino, substituted amino, cycloalkyl, substituted cycloalkyl, aryl,substituted aryl, heterocyclic and substituted heterocyclic; b is 1 or2; and X is hydroxyl; and and enantiomers, diastereomers andpharmaceutically acceptable salts thereof.
 12. The compound of claim 11wherein the compound is selected from formula IIc, IId or IIe.
 13. Thecompound of claim 11 or 12 wherein HetAr is a nitrogen containingheteroaryl group which is substituted with a group of formula—O-Z-NR¹¹R^(11′) or —O-Z-R¹² wherein R¹¹ and R^(11′) are independentlyselected from the group consisting of hydrogen, alkyl, substitutedalkyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, heterocyclic, substituted heterocyclic, and where R¹¹ andR^(11′) are joined to form a heterocycle or a substituted heterocycle,R¹² is selected from the group consisting of heterocycle and substitutedheterocycle, and Z is selected from the group consisting of —C(O)— and—SO₂—.
 14. The compound of claim 13 wherein the nitrogen containingheteroaryl group is substituted with a group of formula—OC(O)NR¹¹R^(11′) wherein R¹¹ and R^(11′) are independently selectedfrom the group consisting of alkyl or R¹¹ and R^(11′) are joined to forma heterocycle or a substituted heterocycle.
 15. The compound of claim 14wherein the nitrogen containing heteroaryl group is substituted with—OC(O)N(CH₃)₂ and is at the para position of the heteroaryl group. 16.The compound of claim 11 or 12 wherein HetAr is a nitrogen containingheteroaryl group which is substituted with an aryl or substituted arylgroup.
 17. A method for treating a disease mediated by VLA-4 in apatient, which method comprises administering a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and atherapeutically effective amount of a compound of claims 1 to
 16. 18. Apharmaceutical composition comprising a pharmaceutically acceptablecarrier and a therapeutically effective amount of a compound of claims1-16.